Divergent evolution of wild and cultivated subspecies of <Emphasis Type="Italic">Triticum timopheevii</Emphasis> as revealed by the study of <Emphasis Type="Italic">PolA1</Emphasis> gene

Triticum timopheevii (genome symbol AAGG) comprises two subspecies, cultivated ssp. timopheevii, and wild ssp. armeniacum. These two subspecies are considered as allotetraploids of AA genome from Triticum diploid species and SS genome from Aegilops species. The difference in genome symbol (G vs. S) is due to wide genetic variations among four SS genome species, Ae. bicornis, Ae. longissima, Ae. searsii, and Ae. speltoides. In order to study the origin of T. timopheevii, we compared 19th intron (PI19) sequence of the PolA1 gene, encoding the largest subunit of RNA polymerase I. Two different sized DNA fragments containing PI19 sequences (PI19A and PI19G) were amplified both in ssp. timopheevii and ssp. armeniacum. Shorter PI19A (112 bp) sequences of both subspecies were identical to PI19 sequences of two AA species, T. monococcum and T. urartu. Interestingly, the longer PI19G (241–243 bp) sequences of ssp. armeniacum showed more similarity to PI19 sequences of Ae. speltoides whereas ssp. timopheevii showed more similarity to PI19 sequences of other three SS genome species. The results indicated that two subspecies of T. timopheevii, ssp. armeniacum or ssp. timopheevii, might have arisen independently by allotetraploidization of AA genome with Ae. speltoides or one of the remaining three Aegilops species, respectively.     

Characterization of the Wx gene in diploid Aegilops species and its potential use in wheat breeding

Wx gene encodes for the granule-bound starch synthase I or waxy protein, which is the sole enzyme responsible for amylose synthesis in wheat seeds. The Aegilops species, which are related to wheat, could be important sources of variation in this gene. In addition to its role in starch quality, this gene has been used in phylogenetic studies of wheat. The current study evaluated the variability of Wx gene in seven diploid species of Aegilops genus and compared their nucleotide sequences with the wheat homeologous genes. Nineteen new Wx alleles were found in the seven species evaluated. The alleles detected in two species of the Sitopsis section, Ae. searsii and Ae. speltoides, were related to the Wx-B1 gene of wheat. Two more of the Sitopsis species did not appear to be associated with this genome, whereas the remaining species were related to the Wx-D1 gene of wheat. The results showed an important variation of the Wx gene present in the Aegilops genus, and the 19 new Wx alleles detected could enlarge the genetic pool of wheat.     

Phylogenetic relationships among Triticum L. and Aegilops L. species as genome progenitors of bread wheat based on sequence diversity in trnT-F region of chloroplast DNA

Cultivated wheat, (Triticum aestivum L.), is one of the most important food crops in the world. The Aegilops L. genus is frequently utilized by plant breeders for improving the current wheat cultivars due to their close relationships. Therefore, understanding the phylogenetic relationships among the species of these genera is not only valuable for plant taxonomy, but also for plant breeding efforts. The presented phylogenetic analysis was based on the sequences of trnT-F chloroplast DNA containing three non-coding sub-regions. Twelve genotypes belonging to four species of Triticum L. genus and twenty-four genotypes belonging to eight species of Aegilops genus were used in the current study. The results postulated a close genetic relationship between diploid Aegilops species containing the BB genome and polyploid Triticum species. With the exception of Aegilops cylindrica Host (CCDD), all other Aegilops species having the CC genome were alienated from Aegilops speltoides Tausch (BB) and clustered together. These two clusters joined by a third cluster including the AA genome containing diploid Triticum species.     

Molecular diversity of restriction enzyme sites, Indels and upstream open reading frames (uORFs) of 5′ untransalted regions (UTRs) of Waxy genes in Triticum L. and Aegilops L. species

5′ Untransalted regions (UTR) sequences of Waxy genes were amplified from all 81 Triticum L. and Aegilops L. species by PCR with specific primers. It was found that the sequence length at 7D loci was longer than that 7B and 7A. These sequences contained 170 singleton variable sites and 484 polymorphic sites and that the average length of Indels was 8.5?bp. There were abundant regions of restriction enzyme sites and two regions of simple sequence repeat, “GAA” and “CTGA”, in all sequences. A total of 65 uORFs were detected and classed into 37 types, with the variation in uORFs mainly due to single nucleotide polymorphisms (SNPs), and also to the presence of Indels. All sequences in tetraploids and hexaploids could be grouped into Types A, B, I, II, GI and GII based on sequence variation. Restriction enzyme sites, Indel polymorphisms and the classes of uORFs present together indicated that Type I was more similar to Ae. tauschii whereas Type II was more similar to Type B, and both more similar to Ae. longissima; Type A was more similar to Ae. speltoides. Population analysis was performed and Neighbour-joining trees derived from different species, types and accessions further confirmed that the ancestors of T. urartu, Ae. speltoides, Ae. longissima and Ae. tauschii were involved to the evolution of common wheat, and also implied that Ae. longissima might have participated later than Ae. speltoides. 5′ UTRs of Waxy genes in tetraploid and hexaploid species conserved characters from their respective progenitors when compared with diploid. For the first time, we are able to conclude there is abundant variation in SNPs, Indels and uORFs between 5′ UTRs of Waxy genes from different species related to common wheat, and suggest that further research could help to understand Waxy gene function more deeply and hence improve wheat breeding. Our results also show that three hexaploid species in China have unique diversity in the 5′ UTR of their Waxy gene.     

Variation in zinc efficiency among and within Aegilops species

Fifteen accessions of Aegilops tauschii (DD), 10 of Ae. speltoides (SS) and 8 of the tetraploid Aegilops species sharing the U genome were used to study the influence of varied zinc (Zn) supply on development of Zn-deficiency symptoms, and on shoot dry weight and Zn concentration. Plants were grown in a Zn-deficient calcareous soil under greenhouse conditions with (+Zn = 5 mg kg^—1 soil) and without (—Zn) Zn supply. Four accessions of wild tetraploid wheat, Triticum turgidum var. dicoccoides (BBAA), a group known for its high sensitivity to Zn-deficiency, were used in the experiments for comparison. As expected, the accessions of wild T. turgidum var. dicoccoides showed the highest sensitivity to Zn deficiency, and had more severe leaf symptoms of Zn deficiency (whitish-brown necrotic patches). Among the Aegilops species, leaf symptoms of Zn deficiency were, in general, more distinct in Ae. tauschii (DD) and least in Ae. speltoides (SS). Zinc efficiency, expressed as the percentage of shoot dry weight produced under conditions of Zn deficiency compared to Zn supply, averaged, 15% for T. turgidum, 32% for Ae. tauschii, 52% for Ae. speltoides and 61% for the tetraploid Aegilops species carrying the U genome. Differences in Zn efficiency among and within Aegilops species and T. turgidum were significantly correlated with the Zn amount per shoot, but not with the Zn amount per unit dry weight of shoots. The results show that Aegilops species can be exploited as an important genetic source for Zn efficiency genes, particularly Ae. speltoides var. ligustica (SS) and Ae. triuncialis (UUCC). Transfer of these genes to cultivated modern wheat may bring about a greater variation in Zn efficiency in wheat, and facilitate production of Zn-efficient modern wheat cultivars for Zn-deficient soil conditions.     

Screening wheat germplasm for seedling root architectural traits under contrasting water regimes: potential sources of variability for drought adaptation

In this study, we selected 180 accessions from different wild relatives of wheat (Aegilops–Triticum species) and tested them in the presence of a tolerant and a sensitive control variety under well-watered and drought-stressed conditions. The results of analysis of variance for dry masses and all measured root traits showed that water regimes, accession and species main effects were highly significant. Drought stress significantly declined shoot dry mass (59.42%), root length (37.85%) and the total number of branch roots (36.25%), but increased the root-to-shoot ratio (75.00%), specific root length (64.19%) and root tissue density (29.46%). Principal component analysis for 182 individuals and 12 species groups identified two components that explained 75.67 and 82.39% of the total variation in dry mass and root traits under drought-stressed conditions, respectively. Taking together, our results identified 12 accessions with superior tolerance to drought stress. Remarkably, four species of wild relatives – Ae. cylindrica (DC genome), Ae. neglecta (UM genome), Ae. speltoides (B genome) and Ae. tauschii (D genome) – responded well to drought stress. The potential of these species could be used for molecular analysis such as marker assisted selection and gene mapping, ultimately aimed at breeding for root traits with improved adaptation to drought environments.     

Size homoplasy and mutational behavior of chloroplast simple sequence repeats (cpSSRs) inferred from intra- and interspecific variations in four chloroplast regions of diploid and polyploid <Emphasis Type="Italic">Triticum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species

Chloroplast simple sequence repeats (cpSSRs) are widely distributed in the chloroplast genomes of all plant species, and are frequently employed for genotypic and phylogenetic analysis. However, information on intra- and interspecies variation in cpSSRs is lacking. In this study, we sequenced four intergenic (non-coding) chloroplast DNA regions in 57 accessions of 12 tetraploid, and 16 accessions of 4 hexaploid species of Triticum and Aegilops. These sequence data added to our previous data for diploid species in the same chloroplast regions. Intra- and interspecific genetic variation was analyzed for a total of 189 accessions of 13 diploid, 12 tetraploid, and 4 hexaploid species of Triticum and Aegilops, such that all species were represented by multiple accessions. The data were used to infer phylogenetic relationships within and among Triticum and Aegilops species. Based on this robust phylogenetic tree, seven of eight cpSSR loci clearly exhibited “size homoplasy,” referring to the fact that cpSSRs of identical size and DNA sequence can arise even if the alleles are not descended from a common ancestor. These data indicate that cpSSRs should be used with caution in phylogenetic analyzes. Interestingly, as observed from several previous studies, our data also suggest that observed mutation rates may increase significantly when mononucleotide (homopolymer) repeat numbers reach or exceed 9 bp. In the present report, using this sequence data set involving cpSSRs, 81 unique haplotypes among 189 accessions were detected, and five tetraploid Triticum and Aegilops species were successfully identified and genotyped. Our results indicate that combinations of nucleotide substitutions, indels and SSRs of chloroplast nucleotide sequences are available for genotyping at the species accession level.     

Relationship between spike morphology and habitat of four <Emphasis Type="Italic">Aegilops</Emphasis> species of section Sitopsis

This study examines the relationship between spike morphology and natural habitat for 84 accessions of four Aegilops species, belongs to section Sitopsis, Ae. bicornis, Ae. longissima, Ae. searsii, and Ae. sharonensis in genus Aegilops, section Sitopsis, wild relatives of Triticum aestivum L. These species are considered valuable genetic resources for future cultivation and breeding of domesticated wheat. The goals of the study were to: (1) document variation in spike morphology among these four species; (2) examine the relationship between spike morphology and native habitat; (3) document geographical distribution of distinct spike morphology; and (4) examine the relationship between spike morphology and heading time and value for these four species. The results reveal significant differences in spike morphology among species of section Sitopsis. The most noteworthy variation involved the absence/presence of lateral awn, such that species with lateral awn were restricted in coastal, though species without lateral awn were mainly distributed in inland. This suggests that local climate may be a determinant of variation in lateral awn, and that this trait may be subject to convergent evolution. Differences in heading time in sympatric area were also observed. The differences may enhance species divergence and could represent a lead speciation event. The results of this study will facilitate identification of populations or accessions of wild wheat with favorable traits and/or novel adaptive genes.     

Complete chloroplast DNA sequences of Georgian indigenous polyploid wheats (<Emphasis Type="Italic">Triticum</Emphasis> spp.) and B plasmon evolution

Three types of plasmon (A, B and G) are present for genus Triticum. Plasmon B is detected in polyploid species - Triticum turgidum L. and Triticum aestivum L. By now, 21 complete sequences of chloroplast DNA of the genus Triticum is published by different authors. Many inaccuracies can be detected in the sequenced chloroplast DNAs. Therefore, we found it necessary to study of plasmon B evolution to use only those sequences obtained by our method in our laboratory. Complete nucleotide sequences of chloroplast DNA of 11 representatives of Georgian wheat polyploid species were determined. Chloroplast DNA sequencing was performed on an Illumina MiSeq platform. Chloroplast DNA molecules were assembled using the SOAPdenovo computer program. Using T. aestivum L. subsp. macha var. palaeocolchicum as a reference, 5 SNPs were identified in chloroplast DNA of Georgian indigenous polyploid wheats. 38 and 56 bp inversions were observed in paleocolchicum subspecies. The phylogeny tree shows that subspecies macha, durum, carthlicum and palaeocolchicum occupy different positions. According the simplified scheme based on SNP and indel data the ancestral, female parent of all studied polyploid wheats is an unknown X predecesor, from which four lines were formed.     

Characterization of Starch in Aegilops Species

Starch was extracted and cleaned from 99 accessions of 20 species of Aegilops and also from 200 accessions of hexaploid wheat. Amylose content was determined by iodine staining and absorbance at 535 and 620 nm. Particle‐size distribution was determined by laser scattering. The amylose content of the Aegilops accessions did not exceed the extremes found in domesticated wheat. Aegilops species, on the whole, had a lower content of small particles than the hexaploid wheats. There was no correlation between amylose content and particle‐size distribution. Some species of Aegilops may be useful sources of low‐starch B‐type granules for hexaploid wheat, if the trait can be transferred, but they are unlikely to contribute to further variation in amylose content.     

RFLP analysis of Aegilops species belonging to the Sitopsis section

The phylogenetic relationships among theAegilops species belonging to the Sitopsis section were investigated using RFLP (restriction-fragment-length polymorphism) analysis. Twenty-five probes, each of which hybridised to oneor more restriction fragments located in the B-genomechromosomes of cultivated wheats, were used. At least one and in most cases two fragments were located in every B genome chromosome arm. Adendrogram derived from a cluster analysis of the complete RFLP dataset showed a subdivision of the species belonging to the Sitopsis section into one group comprising the species of the Truncata subsection and another group comprised of the species of theEmarginata subsection. Dendrograms also were produced using RFLP data from loci located in different combinations of only three chromosomes, and some of these showed different subdivisions of the species. This demonstrates the importance in obtaining reliable classification data of using probes that detect loci evenly distributed in the genome and located in each chromosomearm.     

Characteristics of the root system in the diploid genome donors of hexaploid wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Wild crop relatives are of considerable interest in plant breeding and significant efforts have been made to transfer their genetic variation into modern crops. Of the three diploid progenitors of bread wheat (Triticum aestivum L.), only Aegilops tauschii Coss. has been explored and exploited and only for some above ground characteristics. The three wild progenitors (Aegilops speltoides Tausch., Triticum urartu Tumanian ex Gandilyan, and Aegilops tauschii) have never been assayed for root traits. Here we report such a root study, and include Triticum monococcum L. subsp. boeoticum (Boiss.) Hayek and T. turgidum L. subsp. dicoccoides (Koern. ex Asch. et Graebn.) Thell. Fifteen accessions were selected from the above wild species and tested in the presence of one bread wheat cultivar Pavon F76. Significant variation was observed between and within the taxa. Of all accessions tested, cv. Pavon F76 had the smallest root system at maturity while A. speltoides had the largest root system. Moreover, Aegilops spp. had larger mean values for root biomass when compared with Triticum spp. These results suggest there is significant unexplored potential for the use of wheat wild relatives in wheat breeding to improve the root system, or to develop synthetic mapping populations to study root traits.     

Evaluation of geographic distribution of high-molecular-weight glutenin subunits (HMW-GS) in wild and cultivated Triticum species

Variation of high-molecular-weight glutenin subunit (HMW-GS) in 632 wild and cultivated Triticum accessions was investigated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A total of 11 alleles of HMW-GS in diploid species, 22 in tetraploid species, and 15 in hexaploid species were detected. Diploid species on Glu-1A locus and tetraploid species Glu-1B locus showed the highest diversity, respectively. Tetraploid species had the highest level of diversity on three Glu-1 loci, followed by hexaploid and diploid, based on Shannon’s information index, Nei’s genetic diversity, and percentage of polymorphic loci. Molecular variance analysis confirmed main variance of HMW-GS within species, regions, and locations, respectively. Variance among species and regions was enhanced gradually with the increase of ploidy. Significant non-random distributions between the phylogenic trees of HMW-GS and the locations of accessions were tested by GenGIS software, indicated that geographic factors played an important role along the different orientations in the spread of Triticum species. We found one original diversified center in diploid what located around Elazig, Malatya, Gaziantep, Urfa, and Kiziltepe in Turkey, and three diversified centers in tetraploid wheat, including Turkey–Armenia–Georgia–Iran, Portugal–Spain, and Ethiopia, respectively, and two diversified adjacent areas between Turkey and Switzerland and around Turkey, Georgia, and Armenia. The original center of diploid species located in southeast Turkey, where the unexpressed 1Ay subunit was mainly distributed in T. urartu, could be one of the candidate regions of polyploidization of Triticum L. The regional distribution of HMW-GS and species also provided geographic evidences for the existence of founder effect on the spread of Triticum species. The present study suggests that integrating genetic diversity with geographic characterization in Triticum could very useful for collection, conservation, and utilization, as well as for research microevolution and domestication.     

Red List assessment of nine <Emphasis Type="Italic">Aegilops</Emphasis> species in Armenia

The aims of this study are to determine the geographical and ecological distribution of nine Aegilops species in Republic of Armenia and to make an assessment of their IUCN Red List status, using the IUCN Red list categories and criteria, in order to develop an in situ conservation strategy for wild relatives of wheat in Armenia. Ecogeographic surveys of nine Aegilops species were undertaken over 2 years in Armenia. They included a herbarium survey followed by extensive ground-truthing field surveys where targeted Aegilops species occur. The study showed that of the nine Aegilops species studied, four are threatened and of these, Ae. mutica and Ae. crassa are critically endangered. The latter species may even be extinct in Armenia. Ae. neglecta and A. biuncialis are endangered. Additional studies are required to assess the threat status of Ae. umbellulata. Ae. columnaris was assessed as near threatened, while the remaining species (Ae. triuncialis, Ae. cylindrica and Ae. tauschii) are of least concern. There has been a dramatic decline in the genetic resources of Aegilops species during recent years in Armenia as a result of adverse human impacts such as expansion of agriculture, urbanization and uncontrolled grazing. Several species, especially Ae. mutica and Ae. crassa, should be prioritized in conservation activities in Armenia. Efforts should be made to conserve genetic diversity of crop wild relative species both in situ and ex situ, bearing in mind that their germplasm carries potentially valuable information (traits) that can improve adaptability and productivity of cultivated wheat varieties.     

Molecular characterization and diversity of puroindoline b-2 variants in cultivated and wild diploid wheat

Cloning and phylogenetic analysis of puroindoline b-2 variants in common wheat (Triticum aestivum L.) and its relatives would advance the understanding of the genetic diversity and evolution of puroindoline b-2 gene in common wheat and its related species. In the present study, common wheat (AABBDD) and four related species, including T. urartu (A^uA^u), Aegilops speltoides (SS), Ae. tauschii (DD), and T. turgidum (AABB) were sampled for the presence of novel alleles at Pinb2v-A1, Pinb2v-B1/Pinb2v-S1 and Pinb2v-D1 loci corresponding to common wheat puroindoline b-2 variants. Nine new alleles were identified at these loci, designated Pinb2v-A1a through Pinb2v-A1c, Pinb2v-S1a through Pinb2v-S1e, and Pinb2v-D1a. Alignment of puroindoline variants or alleles from common wheat and its relatives indicated that all alleles in diploid wheats are attributed to single nucleotide substitution when compared with puroindoline b-2 variants in polyploids. Deduced amino acid sequences showed that all three alleles at Pinb2v-A1 locus and four alleles (Pinb2v-S1a, Pinb2v-S1b, Pinb2v-S1c and Pinb2v-S1e) at the Pinb2v-S1 locus could not be normally translated due to the presence of premature stop codons, whereas Pinb2v-D1a at the Pinb2v-D1 locus and Pinb2v-S1d at the Pinb2v-S1 locus could be normally translated, possibly suggesting that the puroindoline b-2 variant in Ae. tauschii was more highly conserved than those in T. urartu and Ae. speltoides. Meanwhile, puroindoline b-2 variant could be normally translated in all of the durum and common wheat cultivars surveyed. None of the puroindoline b-2 alleles previously identified in durum and common wheat were found in the diploid genome donors examined here, even though a greater diversity of alleles were found in diploid wheat compared to polyploid wheat. These results likely reflect the evolutionary history of tetraploid and hexaploid wheats, although it may be that puroindoline b-2 variant alleles have been selected for stability and functionality in common and durum wheat. This study provides a survey of puroindoline b-2 variants in common wheat and its relatives, and provides useful information for understanding the genetic diversity of puroindoline-like genes and their duplication events in wheat.     

Evolution of tetraploid wheat based on variations in 5′ UTR regions of Ppd-A1: evidence of gene flow between emmer and timopheevi wheat

Previous study showed that tetraploid wheat was divided into two groups (Type AI and Type AII) based on sequences around Ppd-A1 gene (Takenaka and Kawahara in Theor Appl Genet 125(5):999–1014, 2012). That study focused on domesticated emmer wheat and used only 19 wild emmer wheats, so could not be clear the evolutional relationship between Type AI and Type AII. Here, a total of 669 accessions comprising 65 einkorn wheats, 185 wild emmer wheats, 107 hulled emmer wheats, 204 free-threshing (FT) emmer wheats, and 108 timopheevii wheats were studied by PCR assay and DNA sequencing for Type AI/AII. Type AII was an older type than Type AI because all einkorn accessions had Type AII. In wild emmer, Type AI was distributed in the northeast regions of its distribution and Type AII was found to be centered on Israel. A total of 37.4 % of hulled emmer accessions were Type AI, while 92.2 % of FT emmer accessions were Type AI. Differences in the proportion of Type AI/AII in domesticated emmer suggested a strong bottle-neck effect. We also found two MITE-like sequence deletion patterns from a part of Type AII accessions (dic-del and ara-del). Dic-del was found from only Israeli wild emmer accessions and ara-del was found from almost all timopheevii wheat accessions. Only three timopheevii accessions did not have ara-del, and one wild emmer accession and ten hulled emmer accessions had ara-del. These accessions suggested gene flow between emmer and timopheevii wheat.     

Stem and leaf rust resistance in wild relatives of wheat with D genome (Aegilops spp.)

Resistance to stem rust and leaf rust in five D genome species of wheat viz., 267 accessions of Aegilops tauschii Coss., 39 of Ae. cylindrica Host, 17 of Ae. ventricosa Tausch, 4 of Ae. crassa Boiss. and 8 of Ae. juvenalis (Thell.) Eig were evaluated at adult plant stage. Two hundred and thirty nine (90 %) accessions of Ae. tauschii, 30 (77 %) of Ae. cylindrica, 16 (94 %) of Ae. ventricosa, 3 (75 %) of Ae. crassa Boiss. and 5 (62.5 %) of Ae. juvenalis were resistant to stem rust pathotypes prevalent in South India at Wellington under field condition. Invariably, all the accessions of the five species were resistant to leaf rust pathotypes. Quantitative measurement of disease using area under the disease progress curve revealed the slow progress of disease in the resistant accessions compared to susceptible check (Agra Local). Since all the five species have D genome, it could be concluded that the genes present in D genome might play a vital role in leaf rust resistance, but in case of stem rust resistance wide range of differential response was noticed. Among the species evaluated, Ae. tauschii was exploited to a larger extent, followed by Ae. ventricosa and Ae. cylindrica for leaf and stem rust resistance because of the homology of D genome with hexaploid bread wheat. While, Ae. crassa and Ae. juvenalis could not be utilized so far, possibly due to partial homology which makes the transfer of traits difficult. So, these species have considerable potential as a source of rust resistance and may enhance the existing gene pool of resistance to stem and leaf rusts.     

The 5S rRNA gene sequence variation in wheats and some polyploid wheat progenitors (Poaceae: Triticeae)

We have cloned and sequenced 115 repeat units of the 5S rDNA genes and spacers from wheat (Triticum) and the polyploid wheat progenitor, Aegilops, and analyzed them together with sequences available in GenBank® (National Center for Biotechnology Information, NCBI, NLM, NIH, Bethesda, Maryland, USA). We were able to assort the sequences into nine orthologous groups which we call unit classes. The following unit classes were assigned to haplomes, and labeled accordingly: long A1, short A1, short A2, long G1, short G1, long D1, short D1, long S1 and short S1. The AA-genome, DD-genome and SS-genome species were each found to contain a long and a short class. The AAGG-genome species T. timopheevii and the AAA^tA^tGG-genome species T. zhukovskyi, both contain the long A1, long G1 and short G1 unit classes. The AABB-genome species T. turgidum consists of the short A2, a unit class not yet found in T. monococcum, and the long S1 unit class found in the species of Aegilops section Sitopsis. The bread wheat AABBDD-genome contains the long A1, short A2, long D1, long S1 and short G1 unit classes. The presence of the long S1, also demonstrated to occur in both T. turgidum and T. aestivum, supports the hypothesis that the progenitor of the B-haplome in wheat originated in Aegilops section Sitopsis. The presence of the short G1 unit class, i.e. the G-haplome in bread wheat, is unexpected. These new findings are discussed in the light of published findings, especially those relating to 5S DNA loci and evolutionary hypotheses.     

Genetic diversity of the D-genome in <Emphasis Type="Italic">T. aestivum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species using SSR markers

Simple sequence repeats (SSRs), highly dispersed nucleotide sequences in genomes, were used for germplasm analysis and estimation of the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (D^tD^t), Ae. cylindrica (CCD^cD^c) and Ae. crassa (MMD^cr1D^cr1), collected from 13 different sites in Iran. A set of 21 microsatellite primers, from various locations on the seven D-genome chromosomes, revealed a high level of polymorphism. A total of 273 alleles were detected across all four species and the number of alleles per each microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Data obtained in this study supports the view that genetic variability in the D-genome of hexaploid wheat is less than in Ae. tauschii. The highest number of unique alleles was observed within Ae. crassa accessions, indicating this species as a great potential source of novel genes for bread wheat improvement. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

Phylogenetic analysis of complete 5′ external transcribed spacers of the 18S ribosomal RNA genes of diploid Aegilops and related species (Triticeae, Poaceae)

PCR systems were designed to amplify the entire 5 external transcribed spacer (ETS) region of the 18S rRNA gene of all the diploid species of Aegilops and several other taxa closely related to domesticated wheat. Phylogenetic analysis was performed on the complete ETS sequences using the neighbor-joining, maximum parsimony, and maximum likelihood methods. Among the individual taxa studied, speciation in Secale is very recent. In the case of the A genome diploids, the results support the theory that the A genomes of wheat have experienced reticulate evolution owing to introgression. The B and G genomes of tetraploid domesticated wheats form a clade with Ae. speltoides in which the B genome diverged first and the G genome more recently. It was demonstrated that the complete ETS sequences of the Triticeae yield coherent phylogenetic information. The ETS is a useful tool for studying the phylogeny of closely related species.