Aegilops tauschii: Genetic Variation in Iran

All the 79 Aegilops tauschii Coss. accessions of Iranian origin from Prof. Kihara’s collection were analyzed electrophoretically. Of 23 enzyme-encoding loci studied, 11 were polymorphic. In Iran Ae. tauschii is presented by ssp. tauschii and ssp. strangulata which distinctly differ genetically, morphologically and ecologically. Variation patterns of low polymorphic locus Aco2 and highly polymorphic Ep are similar in both subspecies. In contrast, variation of Acph1, Ak, Est2, Est5, Got1, Got2, Got3 and Lap is a set of diverse patterns which markedly differ between subspecies and natural regions also, implying that natural selection is involved.     

Geographic patterns of Got1, Got2, Got3 and Est2 enzyme-encoding genes allelic variation in Aegilops tauschii Coss.

Geographic patterns of Got1, Got2, Got3 and Est2 enzyme-encoding genes allelic variation were investigated among 322 accessions of Aegilops tauschii Coss., 161 accessions of ssp. tauschii and 161 accessions of ssp. strangulata, representing all the species area. It was found that: (1) in the two ecologically different subspecies, ssp. tauschii and ssp. strangulata, the patterns of allelic variation of the four genes differ greatly; (2) the same allozymes have originated several times independently in different Ae. tauschii local populations; (3) allelic variation of Got1, Got3 and Est2 in ssp. strangulata corresponds to climatic conditions. The data obtained reflected that Ae. tauschii has been inhabiting its area from ancient times; and allelic variation patterns of Got1, Got2, Got3, Est2 loci were mostly formed by natural selection. Further investigations of these loci with molecular genetic methods are prospective for understanding the peculiarities of Ae. tauschii evolution.     

Chloroplast DNA non-coding sequences variation in Aegilops tauschii Coss.: evolutionary history of the species

Sequences of four chloroplast DNA non-coding regions, about 3,000?bp in total, were analysed in 112 Aegilops tauschii accessions, 56 of ssp. tauschii and 56 of ssp. strangulata, representing all of the species range. One inversion, 8 insertions/deletions, 18 base pair substitutions and 5 microsatellite loci were found. The data revealed that Ae. tauschii originated in Caucasia. Neither of the two Ae. tauschii subspecies was an ancestor to one another. Aegilops tauschii divided into ssp. tauschii and ssp. strangulata at the very beginning of its existence as a species. Subspecies tauschii was the first to start geographic expansion and relatively rapidly occupied a vast area from Caucasia—eastward up to central Tien Shan and western Himalayas. In contrast to ssp. tauschii, geographic spread of ssp. strangulata was a complicated, multi-stage and slow process. At the beginning of ssp. strangulata evolutionary history its major phylogenetic lineage for a lengthy time span had existed as a small isolated population. Several forms of ssp. strangulata, better adapted to relatively moister and cooler habitats, had originated. Each of these forms has gradually forced out ssp. tauschii from some part of its area in the west, up to central Kopet-Dag.     

Aegilops tauschii Coss.: allelic variation of enzyme-encoding genes and ecological differentiation of the species

Three hundred and seven accessions of Aegilops tauschii Coss., including 160 of subsp. tauschii and 147 of subsp. strangulata, representing all the species range—from Turkey to Kirgizstan, were analyzed electrophoretically. Twenty polymorphic enzyme-encoding loci were studied, 10 of which were essentially polymorphic in Ae. tauschii. Climatic data for each of the 307 Ae. tauschii habitats were taken from WORLDCLIM database of computer system ArcGIS. Forty-nine climatic parameters were considered: precipitation, minimum, mean and maximum temperatures for each month, and also the total annual level of precipitation. The data were analyzed with multivariate statistical methods, such as Principal Components Analysis (PCA), Multiple Correspondence Analysis (MCA) and Two-Block Partial Least Squares. Variability of climatic conditions among Ae. tauschii habitats is reflected by the two approximately orthogonal “vectors”. The “first vector” is mostly determined by negative impact of precipitation and minimum temperatures during winter. The “second vector” is mostly determined by negative impact of maximum temperatures during summer, and positive impact of precipitation during late spring and summer. Aegilops tauschii is essentially variable along the “second vector”, and especially high level of variation is characteristic for subsp. tauschii. This variation reflects that Ae. tauschii is very tolerable to the climatic variation during summer season. Aegilops tauschii subsp. strangulata is also characterized by the high level of variation along the “first vector”. Moreover, all the habitats of subsp. strangulata fall into the two distinct separate clusters: the habitats in Precaspian Iran, which have the highest minimum temperatures in winter,—and all the other habitats. In the plot of the first two factors of PCA, the “cluster of Precaspian Iran” can be further divided into “Western Precaspian Iran (WPI)”, having relatively higher level of annual rainfall, and relatively dryer “Eastern Precaspian Iran (EPI)”. This three groups of subsp. strangulata accessions, from WPI, EPI and other areas, are also distinctly differed in enzyme-encoding genes allelic variation, as revealed on the plot of the first two axes of MCA. In contrast to subsp. strangulata, the level of variation of subsp. tauschii along the “first vector” is rather low. It was pointed out that variation along “the first vector” reflects adaptive intraspecies divergence of Ae. tauschii: its subspecies strangulata “prefers” the habitats of seaside climate, with warm and moist winter; while subsp. tauschii mostly occupies the habitats with rather continental climate, with relatively cold and dry winter. Allelic variation of enzyme-encoding genes Acph1, Ak, Est2, Est5, Got1, Got2, and Got3 correlate with climate along “the first vector”. Apparently, polymorphism of these loci were involved into the process of Ae. tauschii intraspecies adaptive divergence. Allelic variation of Cat2 and Fdp loci correspond to climatic variation along “the second vector” in subsp. tauschii. Therefore Cat2 and Fdp are likely to be among the genes which polymorphism “helped” subsp. tauschii to succeed in vast geographical expansion far to the east from Caspian Sea.     

Stripe rust resistance in <Emphasis Type="Italic">Aegilops tauschii</Emphasis> and its genetic analysis

Aegilops tauschii Coss., the D-genome progenitor of common wheat (Triticum aestivum L.) includes two subspecies, tauschii and strangulata (Eig) Tzvel. Subspecies tauschii has a wide geographic distribution spreading westwards to Turkey and eastwards to Afghanistan and China, while ssp. strangulata has a narrower distribution occurring only in two disjoined regions, southeastern Caspian Iran and Transcaucasia. A collection of 56 Ae. tauschii accessions was screened at adult stage against a mixture of pathotypes of stripe rust prevalent in the current wheat production in China. The results for three crop seasons indicated that among the 38 ssp. tauschii accessions, 37 were susceptible and only one was resistant, while all the 18 ssp. strangulata accessions were resistant. These results indicated that stripe rust resistance was related to taxonomic origin. Further genetic analysis revealed the resistance of stripe rust in ssp. strangulata accession AS2388 was conferred by a single dominant gene.     

Distribution and diversity of Aegilops tauschii in Iran

The wide morphological variation of Aegilops tauschii has led to the distinction of different subspecies; a typical ssp. tauschii and a second ssp. strangulata. However some researchers pointed out the existance of the intermediate form among morphologically distinguished subspecies. Distribution, diversity and the relationship between different subspecies and the intermediate form were evaluated in the Iranian Ae. tauschii collection. This collection was classified to 15 different populations according to morphological similarities and the collecting origin of accessions. The highest variation was found in tauschii population of Golestan followed by tauschii populations of Gilan and Ardebill, whereas the lowest variation was observed in tauschii populations of central Iran. Two discriminant functions suggested that the length of rachis node and spikelet glume, particularly, the length/width ratios of these traits had the highest impact on identification of different forms. Mahalanobis distances (D ² ) between the two subspecies along with intermediate form on the multidimensional scaling plot showed that the intermediate form is more similar to ssp. tauschii than ssp. strangulata. Although, the diversity within the ssp. strangulata was not very high, it widely affected the diversity of Iranian accessions of Ae. tauschii through continues crossing with the more diversed subspecies, tauschii, during thousands of years. This fact had lead to expansion of its distribution from its origin to Northern Khorasan, Northern Semnan and Eastern Ardebill by producing the intermediate form.     

Polymorphism of <Emphasis Type="Italic">Got2</Emphasis> DNA sequences sheds light on <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. intraspecies divergence and origin of <Emphasis Type="Italic">Triticum aestivum</Emphasis> L.

DNA sequences of nuclear gene Got2 was studied in 60 accessions of Aegilops tauschii, 29 of subsp. tauschii and 31 of subsp. strangulata. It was found that Got2 allozyme polymorphism in Ae. tauschii is due to a single, unique, mutation which led to replacement of glutamic acid by isoleucine in residue 256 of the enzyme molecule, encoded by Got2. As revealed by Got2 DNA sequences variation, initially in its history Ae. tauschii was presented by subsp. strangulata, and among phylogenetic lineages of subsp. strangulata, the lineage “t-9¹s” (TauL3) is the most ancient, a relict one. Subspecies tauschii is relatively “young”. Initially it was presented by the lineage marked by combination of allozyme alleles Got2 ¹⁰⁵ and Acph1 ¹⁰⁰. In the past it inhabited the Continental area from Caucasia to Pakistan, but later on it was forced out by newly originated, now—a major lineage of subsp. tauschii, marked by Got2 ¹⁰⁰. This lineage extended the Continental area of the species up to Kirgizstan, but actually failed to penetrate into pre-Caspian area, occupied by subsp. strangulata. These results essentially differ from those obtained previously, using chloroplast DNA (cpDNA) sequences polymorphism. As revealed by cpDNA, the major, “usual”, subsp. strangulata (TauL2) is “younger” than subsp. tauschii, which resided on phylogenetic tree between relict lineage “t-9¹s”of subsp. strangulata—and major subsp. strangulata. But both cpDNA and Got2 DNA sequences indicate that the level of genetic variation in subsp. tauschii is much lower than in subsp. strangulata. According to Got2 DNA sequences variation, it was Ae. tauschii subsp. strangulata lineage “k-109″ which donated genome D to Triticum aestivum L. This lineage includes accessions: k-109 from South-Eastern Precaspian Azerbaijan; KU-2105, KU-2159 from Western Precaspian Iran; KU-2080 from Eastern Precaspian Iran.     

Microsatellite analysis of genetic diversity and population genetic structure of <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. in Northern Iran

Genetic diversity and population genetic structure of Aegilops tauschii in Northern Iran were studied based on nine microsatellite loci. A high level of genetic diversity was observed from the accessions collected from six regions (provinces). These accessions include 79 samples of the two subspecies (tauschii and strangulata), the intermediate form (among morphologically distinguished subspecies) and ten accessions of Triticum aestivum. The nine microsatellites revealed a total of 141 alleles, with an average of 15.7 alleles per locus. A comparison of the parameters showing genetic diversity, including the observed heterozygosity (Ho), gene diversity (He) and Shannon’s information index (I) of Ae. tauschii accessions from different provinces in Northern Iran, indicated that subsp. tauschii possesses the highest genetic diversity, followed by intermediate form. Genetic distance between subsp. strangulata and subsp. tauschii was low, confirming high gene flow between these two subspecies. However, intermediate form was more distinct from both of them. It was also found that the genetic diversity of T. aestivum is obviously lower than that of Ae. tauschii accessions. Moreover, the level of genetic diversity for Gilan, Golestan and Mazanderan provinces was higher than for Ardebil, Ghazvin and Semnan provinces, suggesting that these regions may provide a readily available source of potentially useful variation for wheat improvement.     

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.     

Haplotype variations of gene Ppd-D1 in Aegilops tauschii and their implications on wheat origin

The Ppd-D1 controlling photoperiod response is an important gene for wheat adaptation since it affects heading time. In the present study, three haplotypes, i.e. haplotype I without deletion, haplotype II with a 24?bp deletion, and haplotype III with two deletions of 24 and 15?bp, were identified in the upstream of the coding region in 80 Ae. tauschii accessions. The haplotype distribution was related to subspecies taxon. All typical ssp. tauschii accessions had haplotype I, whereas all ssp. strangulata had haplotype III. The three haplotypes were observed in Ae. tauschii with morphologically intermediate forms between the two typical subspecies. Present results supported that ssp. strangulata or intermediate form was the D-genome donor of common wheat since only haplotype III were found in wheat. Moreover, a 16?bp deletion in exon 8 of gene Ppd-D1 exists in common wheat. However, none of Ae. tauschii accessions analyzed had the 16?bp deletion.     

Multivariate analysis of genetic variation in Aegilops tauschii from the world germplasm collection

A study of Aegilops tauschii subspecies constitution was undertaken. The data on allozyme and morphologic variation among 308 plants from 154 accessions were used for multivariate analysis. ACPH1 and (glume width)/(rachis segment width) ratio were found to be reliable criteria to distinguish between sspp. tauschii and strangulata.     

Polymorphism of gliadins in <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. local populations in two primary habitats in Dagestan

Polymorphism of gliadins was investigated in Aegilops tauschii from primary habitats “4”, near Hily, and “6”, near Rukel, in Dagestan, Russia 205 individual plants were analysed (53/50 and 54/48 plants of subsp. tauschii/subsp. strangulata from the habitats “4” and “6”, respectively) and 1/7 and 18/14 different haplotypes were found among the plants of subsp. tauschii/subsp. strangulata from the habitats “4” and “6”, respectively. No direct evidences of cross-pollination were pointed out, although gliadins electrophoretic phenotypes obtained allowed to suggest that it occur in Ae. tauschii with very low frequency. The data obtained revealed that during Ae. tauschii evolutionary history a local habitat could be populated many times by different phylogenetic lineages of the species. It was found that in Dagestan, at the very edge of the species area, several different lineages belonging to different subspecies could for a long time co-exist together in a local habit, and in such case a very high level of genetic variation in Ae. tauschii could be accumulated on a square of less than one hectare. The further studies of genetic variation in Ae. tauschii local populations, based on molecular genetic methods seems to be very prospective for understanding of peculiarities of the species evolution.     

Purification and Characterization of the Glutenin Subunits of Triticum tauschii,Progenitor of the D Genome in Hexaploid Bread Wheat

N-terminal amino acid sequences and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) molecular weights have been determined for high-performance liquid chromatography (HPLC)-purified high molecular weight (HMW) and low molecular weight (LMW) glutenin subunits (GS) of Triticum tauschii ssp. strangulata, contributor of the D genome to hexaploid bread wheat. The use of three different extraction procedures resulted in similar glutenin preparations. On the basis of N-terminal sequences, the same types of glutenin subunits that have been reported in bread and durum wheats (HMW-GS of both the x and y types and LMW-GS of the LMW-s, LMW-m, α-, and γ-types) were found in T. tauschii. However, the HMW-GS in T. tauschii were in greater proportion relative to LMW-GS when compared to reported values for a bread and durum wheat. Our results support the likelihood that differences in the proportions of the various subunits contributed by the A, B, and D genomes, rather than qualitative differences in the types of subunits, are responsible for the major differences in quality characteristics between bread wheat and durum wheat.     

Comparison of the genetic structure of populations of wild emmer wheat, <Emphasis Type="Italic">Triticum turgidum</Emphasis> ssp. <Emphasis Type="Italic">dicoccoides</Emphasis>, from Israel and Turkey revealed by AFLP analysis

The aim of the present study was to assess the genetic variation in several Israeli and Turkish populations of wild emmer wheat, Triticum turgidum ssp. dicoccoides, the progenitor of most domesticated wheat. Single spikes were collected in 2002 from 60 plants that grew in six different habitats in Ammiad, northeastern Israel (8–12 plants from each habitat), and in 1998 from 56 plants that grew in seven different habitats in Diyarbakir, southeastern Turkey (8 plants from each habitat). Seeds were planted in a nursery and DNA was extracted from every plant and analyzed by the fluorescent-based amplified fragment length polymorphism (AFLP) method. Seven primer combinations produced 788 discernible loci of which 48.6% were polymorphic in Israel and 40.5% in Turkey. The genetic diversity estimates P (frequency of polymorphic loci) and He (gene diversity) were higher in Ammiad than in Diyarbakir (means of P = 0.34 and He = 0.13 in Ammiad vs. P = 0.20 and He = 0.08 in Diyarbakir). Ammiad populations contained more unique alleles than Diyarbakir populations. The relative genetic diversity estimates (θ) values were 0.188 in Ammiad and 0.407 in Diyarbakir, suggesting better differentiation of the populations in Turkey. Genetic distance was larger between Israeli and Turkish populations than between populations of each country. The data indicate that the Israeli and Turkish populations are considerably diverged and that the Israeli populations are more polymorphic than the Turkish ones, having a larger within-populations genetic variation than among-populations one. The significance of the results in relation to the differentiation pattern of wild emmer in the Near East is discussed.     

Durum wheat cultivation associated with Aegilops tauschii in northern Iran

Hexaploid bread wheat (Triticum aestivum L. ssp. aestivum) is assumed to have originated by natural hybridization between cultivated tetraploid Triticum turgidum L. and wild diploid Aegilops tauschii Coss. This scenario is broadly accepted, but very little is known about the ecological aspects of bread wheat evolution. In this study, we examined whether T. turgidum cultivation still is associated with weedy Ae. tauschii in today’s Middle Eastern agroecosystems. We surveyed current distributions of T. turgidum and Ae. tauschii in northern Iran and searched for sites where these two species coexist. Ae. tauschii occurred widely in the study area, whereas cultivated T. turgidum had a narrow distribution range. Traditional durum wheat (T. turgidum ssp. durum (Desf.) Husn.) cultivation associated with weedy Ae. tauschii was observed in the Alamut and Deylaman-Barrehsar districts of the central Alborz Mountain region. The results of our field survey showed that the T. turgidum–Ae. tauschii association hypothesized in the theory of bread wheat evolution still exists in the area where bread wheat probably evolved.     

Novel and ancient HMW glutenin genes from Aegilops tauschii and their phylogenetic positions

A pair of novel high-molecular-weight glutenin subunits (HMW-GS) 1Dx3.1^t and 1Dy11*^t were revealed and characterized from Aegilops tauschii Coss. subspecies tauschii accession AS60. SDS-PAGE band of 1Dx3.1^t was between those of 1Dx2 and 1Dx3, while 1Dy11*^t was between 1Dy11 and 1Dy12. The lengths of 1Dx3.1 ^t and 1Dy11* ^t were 2,514?bp and 1,968?bp, encoding 836 and 654 amino acid residues, respectively. Their authenticity was confirmed by successful expression of the coding regions in Escherichia coli. Network analysis indicated that 1Dx3.1 ^t together with other five rare alleles only detected in Asia common wheat populations represented the ancestral sequences in Glu-D1 locus. Neighbor-joining tree analysis of previously cloned x-type and y-type alleles in the Glu-D1 locus supported the hypothesis that more than one Ae. tauschii genotypes were involved in the origin of hexaploid wheat and that different Ae. tauschii accessions contributed the D genome to common wheat and Ae. cylindrical Host, respectively. An Ae. tauschii accession with 1Dx3.1 ^t or a closely related allele probably have involved in the origin of common wheat. Since accession AS60 used in this study belonged to typical ssp. tauschii, present results suggested the possibility that ssp. tauschii was involved in the evolution of common wheat.     

Analysis of genetic diversity and relationships of wild <Emphasis Type="Italic">Guizotia</Emphasis> species from Ethiopia using ISSR markers

Genetic relationships and diversity of 45 Guizotia populations each consisting of ten individuals and belonging to five taxa of the genus Guizotia were analyzed using Inter Simple Sequence Repeat (ISSR) markers. Five ISSR primers generated a total of 145 scorable bands across the 450 individuals used for the study. The percent polymorphic loci for the taxa ranged from 68.2 (G. arborescens) to 88% (G. scabra ssp. schimperi), with G. scabra ssp. scabra, G. zavattarii and G. villosa following G. scabra ssp. schimperi in this order with respect to the abundance of percent polymorphic loci. The Shannon-Weaver diversity indices (H′), for the five taxa also followed a similar pattern, with G. scabra ssp. schimperi exhibiting the highest H′ (0.7373) and G. arborescens the least (0.5791), while H′ for G. scabra ssp. scabra, G. villosa and G. zavattarii were 0.7313, 0.6620 and 0.6564, respectively. The least genetic distance (0.1188) was observed between G. scabra ssp. schimperi and G.villosa, revealing closer genetic relationships of the two species with each other than with the others, and the highest genetic distance (0.2740) was observed between G. scabra ssp. schimperi and G. zavattarii. The unweighted pair group method using the arithmetic average clustering of the five taxa using the standard genetic distances produced two clusters, with G. scabra ssp. schimperi and G. villosa occurring in one cluster and G. scabra ssp. scabra, G. arborescens and G. zavattarii together in the other cluster. The study reveals that G. scabra ssp. schimperi is more closely related to G. villosa than to G. scabra ssp. scabra.     

Genetic diversity and gene-pool subdivisions of diploid D-genome <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. (Poaceae) in Iran as revealed by AFLP

The diploid goatgrass Aegilops tauschii is considered the D-genome donor of bread wheat and has probably a centre of diversity in north of Iran. In order to measure the genetic diversity of and the relationships among different populations, varieties and subspecies belonging to Ae. tauschii in Iran, DNA was extracted from 48 accessions of Ae. tauschii collected across the geographic range of the species in the Country and the genetic diversity was assessed using AFLPs based on eight PstI/MseI +3 primer pairs resulted in 277 bands, 198 of which were polymorphic. High level polymorphism was detected, with an average of polymorphism rate of 0.715; relatively low genetic similarity (0.455) between accessions and significant difference between the lowest (0.179) and the highest genetic similarity (0.817). The Iranian Ae. tauschii populations showed high level of genetic diversity. The populations studied were divided into two groups: one group was mainly representing Northern populations collected from Southern Caspian Sea shore and the other group was mainly representing Northeast and Northwest populations. Based on the results of this study, it can be suggested that Ae. tauschii possesses two separate gene-pools in Iran: Northern and Northeastern–Northwestern. Considering the needs for introducing new characteristics and alleles for wheat improvement purposes, Ae. tauschii Iranian gene-pool is assumed to be of high importance for more investigation in the future.     

Gliadins of Bulgarian durum wheat (<Emphasis Type="Italic">Triticum durum</Emphasis> Desf.) landraces: genetic diversity and geographical distribution

The polymorphism of gliadins was studied in 98 Bulgarian durum wheat (Triticum durum Desf.) landraces and classified according to the existing catalogues of blocks of gliadin components. In total, 31 alleles, including 12 new ones, were revealed for five gliadin-coding loci. Nine allele families, which included several alleles coding similar blocks differed only by minor components, were found. The gliadin loci had a high genetic diversity (H = 0.70), and Gli-A2 ^d was the most polymorphic locus. Significant differences in allele distribution were observed through the Bulgarian region under study. The results made it possible to explain the distribution by historical factors. Presumably, the genetic material flew into the country via two different ways and different durum wheat subgroups contributed to the formation of Bulgarian landraces. The landraces were a result of long-term selection and, probably, had a close association with the history of the human populations of the region.