Spatial and temporal genetic analyses of Ethiopian barley (Hordeum vulgare L.) landraces reveal the absence of a distinct population structure

Ethiopia is known for its wide topographically induced variation that favours a large amount of plant genetic diversity, and determination of the genetic structure of the germplasm is a crucial step towards exploiting this variation. Accordingly, a study was initiated to determine population structure patterns based on spatial and temporal factors and thereby establish group accessions depending on genetic similarity. To achieve this goal, 15 simple sequence repeats (SSRs) were used for 199 barley landraces collected from ten administrative regions of Ethiopia. The analysis of the spatial genetic structure was performed on the entire data set and on two groups created based on the year of collection. The results obtained from the model-based Bayesian clustering revealed 16 unstructured groups. Furthermore, the grouping of the accessions based on SSR markers resulted in hierarchical chain-type clusters with most of the accessions in the first cluster. The spatial correlations between the genetic and geographical distances revealed a weak population structure for the entire data set, and a weak temporal population structure was also observed for the barley populations from the recent collection. In general, the results indicated the absence of a distinct and consistent population structure among the regions, which could be presumed because of high gene flow. These findings will facilitate parent selection in order to broaden the genetic base of modern cultivars via breeding efforts and also provide information for the planning of an efficient germplasm collection strategy.     

Geographic Pattern of Genetic Diversity Among 43 Ethiopian Mustard(Brassica carinata A. Braun) Accessions as Revealed by RAPD Analysis

As an oilseed crop, the cultivation of Ethiopian mustard (Brassica carinata) is restricted only to Ethiopia. Even though geographic diversity is a potent source of allelic diversity, the extent of genetic diversity among germplasm material of Ethiopian mustard from different countries has not been assessed. Forty-three accessions, comprising 29 accessions from eight different geographic regions of Ethiopia and 14 exotic accessions from Australia, Pakistan, Spain, and Zambia were analysed for their genetic diversity using random amplified polymorphic DNA (RAPD) technique. A set of 50 primers yielded a total of 275 polymorphic bands allowing an unequivocal separation of every Ethiopian mustard accession. The usefulness of the 50 RAPD primers in measuring heterozygousity and distinguishing accessions was variable such that polymorphic information content (PIC) varied from 0.05 to 0.40, band informativeness (BI) from 0.05 to 0.65 and primer resolving power (RP) from 0.15 to 6.83. Jaccard's similarity coefficients ranged from 0.44 to 0.87 indicating the presence of a high level of genetic diversity. On the average, Australian and Ethiopian accessions were the most similar while, Spanish and Zambian accessions were the most distant ones. Cluster analysis grouped the 43 accessions into four groups, which has quite a high fit (r = 0.80) to the original similarity matrix. With no prior molecular information, the RAPD technique detected large genetic diversity among the 43 accessions from five different countries and their grouping by dendrogram and principal coordinate analysis (PCoA) was inclined towards geographic differentiation of RAPD markers. Conversely, RAPD differentiation along geographic origin was not apparent within the Ethiopian accessions.     

Genetic differentiation of <Emphasis Type="Italic">Actinidia chinensis</Emphasis> and analysis of gene flow barriers in the Qinling Mountains,the species’ northern distribution boundary

The Qinling Mountains form the geo-ecological boundary between subtropical and warm-temperate zones in China and represent the northern boundary for the range of native Actinidia chinensis. To protect, explore and make use of natural resources more effectively, factors influencing genetic diversity of species must be determined. Whether the complex demography of the Qinling Mountains significantly influences gene flow of Actinidia species remains unknown. We assessed genetic diversity and structure of A. chinensis populations in the Qinling Mountains using genomic- and EST-SSR markers. We also conducted barrier analysis to detect genetic discontinuity and isolated pools within structured populations. A total of 179 alleles were detected in six natural A. chinensis populations with an average of 14.9 alleles per locus. Overall, genetic differentiation among A. chinensis populations was low but was stronger than that of populations on either the northern or southern slopes. Analysis of molecular variance showed that 25.97% of the total variance occurred among populations, whereas 74.03% of the total variance occurred within populations. The strongest gene flow occurred between the PX population on the southern slope and the LX population on the northern slope. A UPGMA dendrogram revealed that LX and PX populations were clustered together in one group. The other populations were clustered into another group. A Mantel test revealed no significant correlation between genetic and geographical distances. Clear genetic structure was revealed, and four geographic barriers were identified within and among five areas located in the western, central and eastern Qinling Mountains. Gene flow among populations was restricted by the primary ridges of the Qinling Mountains extending west to east, with branches advancing north to south.     

Genetic characterization of the orphan crop tef [Eragrostis tef (Zucc.) Trotter] accessions using simple sequence repeat markers

Tef is the most important and well adapted cereal crop in Ethiopia and grows better than other cereals both in biotic and abiotic stress conditions. In this study 93 tef accessions representative of various agro-ecologies of Ethiopia along with four improved varieties, one local check and six wild relatives were studied. The materials were genotyped with 12 simple sequence repeat markers (SSRs) to investigate the extent and structure of the genetic diversity of tef. A total of 152 alleles were detected indicating high polymorphism in the studied accessions. The number of alleles per polymorphic loci ranged from seven for CNLT 538 to 29 for CNLT150 with an average of 12.67. High average polymorphic information content (0.82) and average expected heterozygosity (0.86) also signifies the suitability and efficiency of SSR markers to discriminate among tef genotypes. Cluster, principal coordinate and structure analyses grouped the 98 tef genotypes and six wild relatives into four distinct classes. The groupings showed no special pattern despite the different regions of origins for the accessions; however, the five wild relatives were grouped together. The analysis of molecular variance and fixation index showed low genetic differentiation and high gene flow between regions revealing that much of the variability was attributed to within populations variation. Generally, the present study showed that there is high genetic variability with unstructured genetic pattern among the studied tef genotypes which implies the vitality of conserved tef germplasms at Ethiopian Biodiversity Institute and the impact of the seed exchange system on tef breeding activities in Ethiopia.

     

Biodiversity of <Emphasis Type="Italic">Secale strictum</Emphasis> in Iran measured using microsatellites

The mountain rye Secale strictum is native to the Middle East and is the progenitor of the cultivated rye. Regarding lack of information about the genetic diversity of this species in Iran, this study was aimed to evaluate its genetic variation and to examine the patterns of diversity related to the varieties and geography. Fifteen wheat and rye derived microsatellite markers were used to achieve this aim. High levels of diversity, with an average number of 6.1 alleles per locus (ranging up to 11) and high level polymorphism with polymorphism rate averaging 0.624 (between populations) and 0.357 (within populations) were observed among 125 individuals from 19 populations collected from various regions of Iran. The Northwestern populations showed the highest and the Northern populations showed the lowest polymorphism and diversity. One population of the Northwest of the country was notably closer in its allele range to the S. cereale accessions used as outgroup. No taxon or geographic specific marker was detected, suggesting high gene flow between the populations, however some groupings which can be related to the geographic regions and varieties, were evident. The analysis of molecular variance attributed same portions of genetic diversity to the within and between populations with no significant variation among different geographic regions. The results of this study indicated that the Iranian genepool of Secale strictum is valuable to search for new useful alleles for crop improvement.     

Population genetic structure of in situ wild Sorghum bicolor in its Ethiopian center of origin based on SSR markers

In situ population studies of wild relatives of crops are crucial for the conservation of plant genetic resources, especially in regions with high genetic diversity and a risk of local extinction. Ethiopia is the center of origin for sorghum, yet little is known about the genetic structure of extant wild populations. Using 9 Simple Sequence Repeat loci, we characterized 19 wild populations from five regions, 8 local cultivar populations from three regions, and 10 wild sorghum accessions from several African countries. To our knowledge, this is the most comprehensive study to date of in situ wild sorghum populations in Africa. Genetic diversity corrected for sample size was significantly greater in the wild populations in situ than in local cultivars or the accessions. Approximately 41 % of the genetic variation in the wild plants was partitioned among populations, indicating a high degree of differentiation and potential value for germplasm conservation, and the average number of migrants (N_m) per generation was 0.43. Cluster analyses showed that some wild populations were grouped by geographic region, whereas others were not, presumably due to long-distance seed movement. Four wild populations from disjunct regions formed a unique cluster with an Ethiopian accession of subsp. drummondii and probably represent a weedy race. STRUCTURE and other analyses detected evidence for crop-wild hybridization, consistent with previous molecular marker studies in Kenya, Mali, and Cameroon. In summary, in situ wild sorghum populations in Ethiopia harbor substantial genetic diversity and differentiation, despite their close proximity to conspecific cultivars in this crop/wild/weedy complex.     

Genetic Diversity Among Traditional Ethiopian Highland Maize Accessions Assessed by Simple Sequence Repeat (SSR) Markers

Over the past three centuries, maize has become adapted to complex environmental conditions in the highlands of Ethiopia. We analyzed 62 traditional Ethiopian highland maize accessions, using 20 simple sequence repeat (SSR) markers and 15 morphological traits, to assess genetic diversity and relationships among these accessions and to assess the level of correlation between phenotypic and genetic distances. The accessions varied significantly for all of the measured morphological traits. The average number of alleles per locus was 4.9. Pair-wise genetic dissimilarity coefficients ranged from 0.27 to 0.63 with a mean of 0.49. Ward minimum variance cluster analysis showed that accessions collected from the Northern agroecology were distinct from the Western and Southern agroecologies. However, there was no differentiation between the Western and Southern accessions. This suggested gene flow between these regions. The relationship between morphological and SSR-based distances was significant and positive (r = 0.43, p = 0.001). The high genetic diversity observed among these set of accessions, suggests ample opportunity for the development of improved varieties for different agroecologies of Ethiopia. From conservation perspective, sampling many accessions from all agroecologies would be an effective way of capturing genetic variation for future collections and conservation.     

Habitat fragmentation reduces genetic diversity and connectivity among toad populations in the Brazilian Atlantic Coastal Forest

Tropical rainforests are becoming increasingly fragmented and understanding the genetic consequences of fragmentation is crucial for conservation of their flora and fauna. We examined populations of the toad Rhinella ornata, a species endemic to Atlantic Coastal Forest in Brazil, and compared genetic diversity among small and medium forest fragments that were either isolated or connected to large forest areas by corridors. Genetic differentiation, as measured by F_ST, was not related to geographic distance among study sites and the size of the fragments did not significantly alter patterns of genetic connectivity. However, population genetic diversity was positively related to fragment size, thus haplotype diversity was lowest in the smallest fragments, likely due to decreases in population sizes. Spatial analyses of genetic discontinuities among groups of populations showed a higher proportion of barriers to gene flow among small and medium fragments than between populations in continuous forest. Our results underscore that even species with relatively high dispersal capacities may, over time, suffer the negative genetic effects of fragmentation, possibly leading to reduced fitness of population and cases of localized extinction.     

Inferring geographic origin of barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L. subsp. <Emphasis Type="Italic">vulgare</Emphasis>) accessions using molecular markers

The USDA Agricultural Research Service (ARS) National Small Grains Collection (NSGC) has 207 landrace barleys obtained from a nursery grown in the Ukraine in 1930 by N.I. Vavilov, many of which have multiple resistance (MR) to disease similar to accessions from Ethiopia. The original collection locations of the accessions from the Vavilov nursery are unknown. The purpose of this study was to determine, using molecular markers, if the MR accessions of unknown origin from the Vavilov nursery are genetically related to MR accessions from Ethiopia. The genetic relatedness among susceptible accessions of unknown origin and among a selection of landrace accessions from the NSGC barley core subset was also assessed. Simple sequence repeat (SSR) and Diversity Array Technology PL (DArT) marker data were used to generate similarity matrices with Dice’s similarity coefficients. Cluster analysis from these results showed that the unknown-origin MR accessions grouped with accessions from Ethiopia. Susceptible accessions of unknown origin were genetically similar to accessions from western Asia, especially near the Caucasus, and from both southern and northern Europe. Based on similarity of marker profiles, some accessions from the core collection are likely duplicates. Future work will seek to identify the probable origin of the remainder of the unknown origin accessions and to more thoroughly characterize the genetic diversity within NSGC barley core subset.     

Genetic differentiation and geographical distribution of barley germplasm based on RAPD markers

Random amplified polymorphic DNA (RAPD) analysis was used to characterize barley germplasm genetic diversity. For the analysis 303 morphologically distinctive accessions were selected from the VIR germplasm collection, St. Petersburg, Russia and the MAFF Genebank, Tsukuba, Japan to represent the principal regions of barley cultivation. A total of 93 polymorphic bands scored from RAPD patterns were used to generate a genetic distance matrix, which was used in both cluster and principal coordinate analysis. Both analysis clearly separated barley cultivars and local populations into three distinctive groups, which evidently reflect different directions in evolution and geographical distribution of barley. The hierarchy of accessions clustering in the first group indicates the westward distribution of barley from West Asia to Europe and New World across Ethiopia and then Mediterranean region. The principal breeding trends based on spike morphology are also observed in this group. The second group is associated with eastward distribution of the crop and represents a unified genetic group, which consists of East Asian and Central Asian accessions. The third distinctive group identified is connected with the evolution and dissemination of hulless forms in Central Asia and the Caucasus region. The conformity of identified genetic groups and clusters with the global centers of crops diversity (gene centers) determined by Vavilov (1926) and modern ecogeographical classification of barley is discussed.     

Whole-genome analysis with SNPs from BOPA1 shows clearly defined groupings of Western Mediterranean, Ethiopian, and Fertile Crescent barleys

The discovery of Hordeum spontaneum C. Koch, a wild ancestor of cultivated barley, in Morocco in 1978 led to the proposal of a multicentric origin for this crop, as an alternative to the widely accepted theory of a single centre of domestication in the Fertile Crescent. Since this discovery, we have tested this hypothesis using the most advanced genetic techniques available at the time, from CM-proteins to RFLP and DNA-chloroplast markers. Nowadays, the availability of single nucleotide polymorphism (SNP) markers that are spread densely over the barley genome provides us with another powerful tool to give further support for the above. We have used 1,536 SNPs from the Barley Oligo Pool Assay 1 (BOPA1) of Illumina to characterize 107 wild and cultivated barley accessions from the Western Mediterranean, Fertile Crescent, Ethiopia, and Tibet. The results have confirmed that each location of the above-mentioned germplasm groups clusters separately. Analysis of molecular variance enabled us to focus on the chromosomal regions and loci that differentiated these groups of barley germplasm. Some of these regions contain vernalization and photoperiod response genes, some of the so-called domestication genes, as well as the most important quantitative trait locus for flowering time in the Mediterranean region. We have combined these results with other genetic evidence, and interpreted them in the framework of current theories on the onset of the Neolithic revolution in the Mediterranean region, to conclude that neither Ethiopia nor the Western Mediterranean can be ruled out as centres of barley domestication, together with the Fertile Crescent.     

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.     

Spatial Distribution of Genetic Diversity in Wild Populations of Phaseolus vulgarisL. from Guanajuato and Michoacán, Méexico

The diversity, genetic structure, and genetic flow of wild populations of Phaseolus vulgaris L. within its Mesoamerican area of domestication, were analyzed by means of morphological and inter-simple sequence repeat molecular markers. Overall, 89% of the loci studied were polymorphic, 35% in the least diverse population and 65% in the most diverse. Genetic diversity in the populations was high, between h = 0.14 and 0.29, as was the maximum distance between populations (D = 0.3). Between 40% and 45% of the diversity was explained by the differences among populations, indicating that a large number of populations is necessary to represent the wild gene pool in the germplasm collections. We found uniformity in allele frequencies among the populations, suggesting presence of outcrossing. We did not find correlation between genetic and geographic distances, but the dendrogram topology suggests geographical isolation due to the mountainous topography. Negative correlations were observed between the coefficient of variation of seed size and the distance between wild populations and fields . We obtained a highly negative correlation between percentage of polymorphic loci and distance to the nearest crop field, which also suggests gene flow from the domesticated populations. These observations suggest that genetic flow is taking place from domesticated toward wild populations and that the farmer, through his agricultural activities, could be influencing the magnitude and the characteristics of the gene flow, and along with this, the differentiation of wild populations. New approaches should be established for conservation in situ and maintaining bio-safety, given the risk of introducing genotypes from the Andes and transgenic varieties and causing genetic assimilation.     

Inferring seed exchange between farmers from population genetic structure of barley landrace Arabi Aswad from Northern Syria

Apart from their significance in genetic conservation, barley landraces are still favoured by local farmers in low-input farming systems. They often perform more predictably under adverse conditions than modern cultivars due to local adaptation. Increased seed exchange between farmers may put adaptation of local populations of barley landraces at risk. Isozyme markers were used to investigate differentiation and infer gene flow between local populations of barley landrace Arabi Aswad in Northern Syria. Inferred gene flow decreased exponentially with geographic distance and may imply that seed exchange between farmers is limited to a regional scale and diminishes over longer distance. Gene flow seems to be a suitable index for estimating seed exchange in highly inbreeding crop species such as barley. In the future, improved mobility of farmers and extension work may facilitate seed exchange over longer distances, and consequently jeopardise preservation of locally adapted landraces in barley.     

Evidence for two domestication events of hyacinth bean (<Emphasis Type="Italic">Lablab purpureus</Emphasis> (L.) Sweet): a comparative analysis of population genetic data

Studying molecular genetic relationships can substantially contribute to the understanding of the pathways of domestication of a species. Although an increasing number of molecular genetic studies have been performed on Lablab purpureus (hyacinth bean), many covered germplasm of restricted geographic origin or limited intra-specific systematic position. Integrating the molecular diversity found with phenotypic or morpho-agronomic diversity is also deficient. This investigation combines findings of eight molecular genetic studies that include about 400 accessions of both wild and cultivated germplasm, thus providing the largest assessment of diversity in Lablab purpureus to date. In particular, results from a recent molecular investigation (Robotham and Chapman 2015) are revisited and reinterpreted by integrating them with known phenotypic diversity. Wild accessions clearly fall into two types, with characteristic pods—2-seeded and 4-seeded. The large majority of cultivated types are more closely related to 4-seeded pod-types. Certain cultivated 2-seeded pod-type accessions from Ethiopia are genetically closer to wild 2-seeded pod-types. These two major phenotypes are reflected in two chloroplast DNA haplotypes A and B. Hence, two domestication events appear to exist in L. purpureus based on this combined data. No other geographic patterns of diversity, which might assist to trace the dispersal of L. purpureus, were found as cultivated accessions predominantly fell into 2-3 major groups. In all studies, the greatest genetic diversity was found in Africa, making Ethiopia one of the probable centers of domestication.     

Geographic dispersion and varietal diversity in Vicia faba L.

Summary Twenty-six faba bean (Vicia faba L.) populations, from five geographic areas and five varietal types have been assayed electrophoretically for variation in seven isozyme systems. Isozyme frequencies in populations, botanical varieties and geographical areas were submitted to the principal component analysis (PCA). Varieties grouped together showed no differences among them except for isolated populations. Populations from Ethiopia and China lay far apart in PCA from those of other origins. It seems that no genetic divergence at the isozyme level took place during the varietal formation and dispersion process, except when geographical isolation of the population occurred. The results support the hypothesis of an isolation of the crop in Ethiopia and in China.     

A case study of seed exchange networks and gene flow for barley (Hordeum vulgare subsp. vulgare) in Morocco

Local patterns of seed regeneration and trade that occur outside the formal breeding sector (seed exchange networks) can have a strong influence on the genetic diversity and evolution of traditional crop varieties. Despite this, little is known about the extent to which seed exchange networks influence gene flow and genetic structure in traditional crop varieties. Here we study barley (Hordeum vulgare subsp. vulgare) in rural communes of Northern Morocco in 2008 and 2009. We quantified seed regeneration and exchange by farmers within the seed exchange network using structured interviews. Using SSR markers, we also quantified the neutral genetic diversity and structure of a complex of traditional varieties referred to as Beldi that is managed in this exchange network. The majority of farmers (>88 %) report cultivating Beldi. Most seeds of Beldi (70–90 %) are maintained on-farm, while the remainder of seeds are obtained from local markets within the commune. Beldi has high genetic diversity and there is weak but significant genetic structure between communes (F_ST = 0.031). From SSR marker data there is evidence of a high level of gene flow between communes not reported in interviews. Seeds purchased in local markets likely represent seeds from a larger geographic region, leading to lower genetic structure among communes than expected based on the reported level of on-farm seed regeneration and local sourcing of seed. We discuss the implications of this seed exchange network for the conservation of traditional barley varieties in the study region.     

Geographical, altitude and agro-ecological differentiation of isozyme and hordein genotypes of landrace barleys from Ethiopia: implications to germplasm conservation

An analysis of the variability of genes encoding six isozyme systems (15 loci) and two storage proteins (2 loci) in landrace barley from Ethiopia is reported. The materials consisted of populations collected from sites as low as 1650 and as high as 3750 meters, covering a wide range of agro-ecological conditions and geographical areas. Of the 17 loci 7 were polymorphic and 10 monomorphic when the 95% criterion of polymorphism was applied. Despite the disproportionate monomorphic loci, polymorphism was detected in all populations when this criterion is used. The populations were found to possess fairly low mean number of alleles per locus (A = 1.5), low mean value of expected heterozygosity (H = 0.134) and a fairly high mean percentage of polymorphic loci (P = 35.3%). The mean F_ST= 0.474 for the populations is typical of inbreeding species. The result indicated that allelic richness is concentrated in altitude class 3 (2500–3000 m) followed by altitude class 1 (<2000 m). Altitude class 2 (2000–2500) holds an intermediate place though it is the highest in terms of expected heterozygosity (H = 0.245). Higher genetic diversity is concentrated in some geographical regions such as Shewa, Arsi, Bale compared to others (Welo, Gamu Gofa, Gojam). Genetic differentiation among the agro-ecological zones was more profound than both among the altitudes and among regions. Correlation analysis between phenotypic diversity (Shannon-Weaver diversity index) and expected heterozygosity (H) for isozyme/hordein loci revealed non significant associations except with respect to agro-ecological zones. In general, it was detected that sites in highland areas in central and northern regions may be more desirable for in situ conservation than sites in peripheral regions in terms of isozyme/hordein diversity and current rate of varietal replacement.     

Allelic diversity of high molecular weight glutenin subunits (HMW-GS) in Iranian Aegilops tauschii Coss. accessions by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE)

Variation of high molecular weight glutenin subunits (HMW-GS) in 28 Iranian Aegilops tauschii (2n = 2x = 14, DD) accessions studied by sodium dodecyl sulphate electrophoresis method (SDS-PAGE). The results showed high variation of HMW-GS in the accessions. The range of frequency in 14 HMW-GS combinations was 3.57–25 % in the accessions. AMOVA showed the molecular variance between the geographic areas was lower than within the geographic areas. According to Nei’s genetic diversity, the highest diversity levels were in Semnan, Golestan and Azarbayjan, on the other hand the lowest levels of diversity were found in Khorasan, Gilan and Mazandaran accessions. Hence, the Caspian Sea South East accessions also Azerbayjan in Iran have more diversity. AMOVA did not show variance between strangulata and tauschii but there was more genetic diversity in ssp. tauschii subspecies in comparison of ssp. strangulata according to Nei’s gene diversity and Shannon information index. It showed Iranian Ae. tauschii have a good potential for bread making quality improvement in bread wheat.     

Genetic Diversity in Tef [<Emphasis Type="Italic">Eragrostis tef</Emphasis> (Zucc.) Trotter] Germplasm

One hundred and forty-four heterogeneous tef germplasm accessions collected from 10 major tef growing areas in Ethiopia were evaluated for 18 quantitative traits in a simple lattice design, at Holetta and Ginchi, in 2001 main growing season to study the genetic diversity in tef. The combined analysis of variance revealed the presence of significant variation among germplasm accessions for all traits studied. Cluster analysis revealed the overlapping of the germplasm accessions from different origin and the accessions were grouped into eight distinct clusters of 1–78. Generalized distance square confirmed the presence of significant genetic distance between clusters. In principal component (Prin) analysis, the first four principal components with latent root values above one accounted for about 80.6% of the variability existing among the germplasm accessions. Prin1 constituted about 55% of the variability mainly from almost equal contribution of 10 quantitative traits, indicating that most of the traits are equally important in tef diversity. In this study, the regions and altitudes of origin were not found to have a substantial effect on the genetic diversity in tef germplasm. Diversity within the regions was found to be significant and, hence, an opportunity for exploitation in tef improvement.