Analysis of genetic diversity and evolutionary relationships among hexaploid wheats <Emphasis Type="Italic">Triticum</Emphasis> L. using LTR-retrotransposon-based molecular markers

The origin, diversity and distribution of hexaploid wheat still remain somewhat unclear. In this study we examined the patterns of genetic diversity and phylogenetic relationships of seven hexaploid wheat species using integration site polymorphism of the LTR retrotransposons. Forty-eight accessions (most of them aboriginal) of seven wheat species from different geographical regions were studied using sequence-specific amplification polymorphisms. Phylogenetic relationships among species were constructed with SplitsTree 4.10 based on Dice’s matrices. Genetic distances between the accessions clustered with PAST software were estimated by principal component analysis. All the accessions differentiated into two main groups, one including European spelt and the other combining common, club and Indian dwarf (shot) wheat with the Asian spelt. The spelt species T. macha, T. vavilovii and spelt spike (speltoid) free-threshing T. petropavlovskyi were intermediate between the two groups. The separation of these spelt species from all other accessions was determined by differences in the A genome. European spelt was subdivided into Central European and Spanish branches. As different genetic pools were characteristic of European and Asian spelt, European spelt could not originate directly from the Asian one. Supposedly, the A genome mostly harbors the species-forming or taxonomically important genes that distinguish spelt species from free-threshing ones, which group together with Asian spelt. Grouping of Asian spelt with free-threshing wheat suggests their close relatedness and confirms the hypothesis that free-threshing hexaploid wheats originated from the Asian spelt ancestor.     

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.     

Genetic relationship of diploid wheat (<Emphasis Type="Italic">Triticum</Emphasis> spp.) species assessed by SSR markers

Genetic diversity of 139 accessions of diploid Triticum species including Triticum urartu, Triticum boeoticum and Triticum monococcum was studied using 11 SSR (simple sequence repeats) markers. A total of 111 alleles with an average of 10 alleles per locus were detected. The polymorphism information content (PIC) of each SSR marker ranged from 0.30 to 0.90 with an average value of 0.62. Among the three Triticum species T. urartu had the highest number of total alleles (Na?=?81), private alleles (Npa?=?15) and showed higher genetic diversity (Hex?=?0.58; PIC?=?0.54). The genotypes from Turkey exhibited the highest genetic diversity (PIC?=?0.6), while the least diversity was observed among 4 Georgian accessions (PIC?=?0.11). Cluster analysis was able to distinguish 139 wheat accessions at the species level. The highest genetic similarity (GS) was noted between T. boeticum and T. monococcum (GS?=?0.84), and the lowest between T. urartu and T. monococcum (GS?=?0.46). The grouping pattern of the PCoA analysis corresponded with cluster analysis. No significant differences were found in clustering of T. urartu and T. monococcum accessions with respect to their geographic regions, while within T. boeoticum species, accessions from Iran were somewhat associated with their geographical origin and clustered as a close and separate group. The results from our study demonstrated that SSR markers were good enough for further genetic diversity analysis in einkorn wheat species.     

Genetic variation for phenolic acids concentration and composition in a tetraploid wheat (<Emphasis Type="Italic">Triticum turgidum</Emphasis> L.) collection

Phenolic acid intake through the consumption of whole-wheat foods provides important health benefits associated with reduced risks of cardiovascular diseases and colon cancer. The genetic variation for phenolic acids was extensively studied in common wheat, but a comprehensive survey in tetraploid wheat is lacking. In this study we evaluated the genetic variability for individual and total phenolic acids concentration existing in a large collection of tetraploid wheat (Triticum turgidum L.). A 2-year evaluation was undertaken on the whole-meal flour of 111 genotypes belonging to seven T. turgidum subspecies including cultivars, landraces and wild accessions. Durum cultivars [T. turgidum subsp. durum (Desf.) MacKey], had the highest average concentration of total phenolic acids (828.7 μg g^?1 dm in 2012; 834.5 μg g^?1 dm in 2013) with amounts varying from 550.9 μg g^?1 dm to 1701.2 μg g^?1 dm, indicating a variation of greater than threefold fold. The lowest concentration of phenolic acids was found in T. turgidum subsp. dicoccum (Schrank ex Schübler) Thell. Rivet wheat (T. turgidum L. subsp. turgidum) had phenolic acid concentrations similar to those in durum, but less variation was noted among the accessions. On the other hand, the accessions of the four remaining subspecies showed lower phenolic acid concentrations and variation among the accessions as compared to durum. A total of six phenolic acids were identified across the wheat genotypes. The effects of genotype, year and year × genotype were estimated by ANOVA and resulted significant for all phenolic acids. The ratio of genotypic variance to total variance suggested the possibility of improving phenolic acid content in elite wheat germplasm through appropriate breeding programs. Moreover, significant correlations between phenolic acids and other quality characteristics of the grain were detected.     

Molecular characterization of <Emphasis Type="Italic">Triticum militinae</Emphasis> derived introgression lines carrying leaf rust resistance

Triticum militinae Zhuk. et Migusch. belongs to timopheevii [Triticum timopheevii (Zhuk.) Zhuk.] group of wheats with 2n = 4x = 28 chromosomes and genome formula A^tA^tGG. Triticum militinae Zhuk. et Migusch. is known to carry resistance to fungal diseases including rusts and powdery mildew. Genes from timopheevii wheat can be incorporated into cultivated wheat by either direct hybridization or through development of amphiploids. Three T. militinae derived introgression lines (ILs) Triticum Militinae Derivative (TMD) 6-4, TMD7-5 and TMD11-5 were selected for the current study based on cytological stability. All three ILs showed resistance against wide spectrum of Indian pathotypes of leaf rust. More than 1200 SSR markers were used for genotyping of ILs and parental lines. The ILs showed variable and multiple introgressions in different chromosomes of A, B and D genome of wheat. The introgression points were distributed mostly in the distal regions though significant introgressions were also observed in proximal regions of some chromosomes. The extent of introgression in ILs TMD6-4, TMD7-5 and TMD11-5 was 2.8, 8.3 and 8.6% respectively. The set of ‘informative markers’ in the Molecularly Tagged Chromosome Regions (MTCR) of T. militinae origin can also be used in future for tagging of genes associated with traits of economic importance apart from leaf rust resistance. The transferability of Triticum aestivum L. SSR markers to T. militinae was 96.4% for A genome, 95.8% for B genome and 84.3% for D genome. Transferability of wheat SSR markers to T. militinae can be used in preparing genetic maps in timopheevii group of wheats.     

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.     

The third glume phenotype is associated with rachilla branching in the spikes of tetraploid wheat (<Emphasis Type="Italic">Triticum</Emphasis> L.)

We investigated the new inflorescence architectural character “three glumes” and a new status of ramification in tetraploid wheat. Using 9 tetraploid accessions with “turgidum type of branching”, the segregation of branched spike and the third glume indicated the complete linkage of genes for both phenotypes. The rachilla were elongated and bore florets at basal nodes while bearing spikelets at the apical nodes in T. durum Desf. TRI 9644 and T. turgidum L. PI 67339. The phenotype was different from “sham-ramification” in Triticum jakubzineri Udacz. et Schachm., with florets at basal and apical nodes of elongated spikelet rachilla. We confirmed a new status of ramification, “false-true ramification”. It was considered that the “false-true ramification” was determined by the shr2 gene in the long arm of chromosome 2A because the ramification of PI 67339 and TRI 9644 was supposed to be allelic. The segregation of “sham-ramification” and the third glume also indicated the complete linkage of genes for both phenotypes. Thus we concluded that the presence of third glume phenotype is associated with rachis and rachilla branching in the spikes of tetraploid wheat. The present study confirmed the existence of three distinct types of spike ramification, whose classification is not entirely unified: (a) “true ramification”—“branched spike”—“genuine branching”—“turgidum type of branching”, (b) “false ramification”—“pseudo-branched spike”—“sham ramification”—“vavilovii type of branching” and (c) “false-true ramification”.     

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.     

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.     

Genetic diversity in tolerance of wild <Emphasis Type="Italic">Avena</Emphasis> species to aluminium (Al)

An evaluation of diversity of aluminium (Al) tolerance of 180 genebank accessions of diploid, tetraploid and hexaploid of wild Avena species from the world collection of the N.I. Vavilov Institute of Plant Genetic Resources (VIR) showed that the accessions with a high degree of aluminium tolerance belonged to the diploids A. canariensis, A. longiglumis and A. wiestii, the tetraploids A. barbata, A. vaviloviana, and hexaploids A. ludoviciana and A. sterilis. A comparison of the data on Al tolerance with the soil conditions demonstrated that most highly tolerance accessions tend to be collected on different type of soils. According to the results of the principal component analysis, preliminary screening for Al tolerance can be carried out among hexaploid species with higher degree of plant resistance to pathogens.     

New sources of compact spike morphology determined by the genes on chromosome 5A in hexaploid wheat

Little is known about the relationship between compact spike loci in hexaploid wheat species. We studied two new compact spike mutants of common wheat Triticum aestivum L. (2n?=?6x?=?42, genome formula BBAADD). The new compact spike genes, C ⁷³⁹ of MCK 739 and Cp of near-isogenic line Mironovskaya 808 (Vrn1), were mapped using aneuploid stocks and microsatellite markers. The C ⁷³⁹ and Cp loci were distally linked with the microsatellite marker Xbarc319 in the F₂ populations of MCK 739?×?‘Novosibirskaya 67’ and Cp-Mironovskaya 808 (Vrn1)?×?‘Saratovskaya 29’. It was evident that the loci affecting compact spikes in T. aestivum mutants were located on chromosome 5AL distal from Q locus. These loci also affected to semi-dwarfism. We named this locus Cp1 (C ompact p lant 1) for all accessions. Cp1 was allelic to C ¹⁷⁶⁴⁸ gene located on the chromosome 5AL of tetraploid wheat [Triticum durum Desf. (2n?=?4x?=?28, genome formula BBAA)]. These dominant genes on chromosome 5AL will be utilized as new gene resources of compact spike morphology in hexaploid wheat. Relationship between loci Q and Cp1 was also discussed.     

Allopatric distributions of tetraploid and hexaploid forms of Aegilops neglecta Req. ex Bertol.

The genus Aegilops L. comprises 22 annual wild species that are closely related to wheat (Hammer in Kulturpflanze 28: 33–180, 1980). Aegilops neglecta Req. ex Bertol. is a member of the Aegilops section of this genus and is distributed from Morocco and Spain in the west to Transcaucasia and western Iran in the east. This species includes tetraploid (2n = 28, genome UUMM) and hexaploid forms (2n = 42, UUMMNN). However, the geographical distributions of the two cytological forms remain unclear. Clarifying the distribution of the two cytological forms is essential for a better understanding of the diffusion of Ae. neglecta and its tetraploid and hexaploid forms. In the present study, chromosome numbers were determined for accessions of Ae. neglecta from a total of 137 populations, located in the western area of the species distribution from the Aegean Islands to Morocco. Taken together with previous studies, the present data reveal a difference in the geographical distribution of tetraploid and hexaploid forms: tetraploid form is distributed in the eastern part of the species area and hexaploid form predominantly occurs in the western part with their border on the western margin of the Aegean Sea. Near the border, tetraploid and mixed populations are sporadically found among hexaploid populations in the Balkan and Peloponnesus Peninsulas, while a few hexaploid and mixed populations are found among tetraploid populations in the East Aegean Islands and West Anatolia.     

Molecular cytogenetic identification of newly synthetic <Emphasis Type="Italic">Triticum kiharae</Emphasis> with high resistance to stripe rust

Six new amphiploids, Triticum kiharae Dorof. et Migusch. (2n?=?6x?=?42, A^tA^tGGDD), are described in this study. They were developed by the chromosome doubling of F₁ hybrid crosses between Triticum timopheevii Zhuk. (A^tA^tGG) with high resistance to stripe rust and Aegilops tauschii Cosson (DD) by colchicine treatment. These amphiploids showed a high level of fertility of 68–80% and exhibited relatively normal chromosome pairing in meiotic metaphase I. Individual chromosomes of T. kiharae could be identified by multicolor fluorescence in situ hybridization using the combination of oligonucleotides probes Oligo-pSc119.2-1, Oligo-pTa535-1, and Oligo-pTa71-2. T. kiharae exhibited high resistance to predominant stripe rust races CYR34, CYR31, CYR32, CYR33, and SY11-4 both during the seedling and adult stages. However, high molecular weight glutenin subunits from Ae. tauschii parents were only partially expressed in the T. kiharae background. These T. kiharae lines provide novel materials to widen the genetic diversity of the common wheat gene pool.     

Microsatellite mapping of the <Emphasis Type="Italic">Scr1</Emphasis> locus conferring screwed spike rachis to modify the spatial orientation of spikelets in <Emphasis Type="Italic">Triticum aestivum</Emphasis> L.

In the wheat (Triticum aestivum L.) inflorescence, spikelets are arranged in two opposite rows on the main axis. Spikelet primordia initiate alternately on opposite sides at angles of 180°. Genotypes exhibiting screwed spike rachis (SCR) have been selected as a gene resource having non-standard spike morphology in wheat. Although the SCR phenotype is a prospective gene resource, it is under-utilized. The SCR phenotype is due to the attachment of spikelets to the rachis nodes on the SCR. Seed number and individual kernel size are critical economic parameters and increasing seed number and single grain weight causes competition among the growing seeds. The SCR phenotype is hypothesized to avoid competition by assuring kernel growth space in each floret. The SCR trait has been observed in spikes and peduncles of KM 60-96. The semi-dwarfism of KM 60-96 was GA₃-sensitive, and it was determined by the presence of Rht8 gene. The response of KM 60-96 to microtubule depolymerizing and stabilizing drugs indicated that the SCR phenotype was not caused by a defect in the α-Tubulin gene. The F₂ of two hexaploid hybrids and a pentaploid hybrid between SCR/normal types segregated 3 SCR:1 normal indicating that the SCR phenotype was determined by a single dominant gene, Scr1. Analysis with microsatellite markers indicated that the Scr1 allele was located in the region between markers Xgwm191 and Xgwm371 in chromosome arm 5BL. From the observation of the backcross generation, introgression of the Scr1 allele into locally adapted wheat cultivars is feasible to increase kernel growth space in each spikelet in the limited spike length.     

Evaluation of genetic resources in the genus <Emphasis Type="Italic">Asparagus</Emphasis> for resistance to <Emphasis Type="Italic">Asparagus virus 1</Emphasis> (AV-1)

Forty-four Asparagus officinalis cultivars, gene bank accessions and breeding lines as well as thirty-four accessions of wild relatives of Asparagus were evaluated for resistance to Asparagus virus 1. Three different test strategies were developed for the assessment of individual plants: (1) natural infection under field conditions, or two vector-mediated infection assays using the green peach aphid Myzus persicae (2) in an insect-proof gauze cage or (3) in a climate chamber. The AV-1 infections were verified by DAS-ELISA and RT-PCR approaches. All tested 660 individual plants of A. officinalis germplasm were susceptible to AV-1 infection. In contrast, in 276 plants of 29 Asparagus wild accessions no virus infection could be detected. These resistant accessions comprised of nineteen diploid, tetraploid and hexaploid species of both the Eurasian clade and the African clade of the asparagus germplasm. Data of the AV-1 resistance evaluation are discussed in relation to the genetic distance of the resistance carrier and potential application in breeding.     

Qualitative traits of perennial wheat lines derived from different Thinopyrum species

Four perennial wheat genotypes derived from crosses between Triticum aestivum and Thinopyrum elongatum, Th. intermedium or Th. ponticum were grown in Central Italy over 2 years of testing, and compared for their agronomical, biochemical, nutritional and technological traits with three commercial common wheat cultivars. Perennial wheat derivatives were characterized by post-harvest regrowth, small kernels, high number of tillers, high protein content and reduced sodium dodecyl sulphate sedimentation volume. Lines 11955 and OK72 exhibited soft kernel texture due to wild-type alleles at the puroindoline loci, whereas lines 235A and 280B produced medium-hard kernels for the presence of novel puroindolines A and B inherited from Th.elongatum and Th. intermedium, respectively. In addition, perennial wheat genotypes presented a high content of carotenoids and 5-n-alkylresorcinols compared with their annual counterparts. AR composition of line 235A, as determined by gas chromatography-mass spectrometry, was characterized by a high percentage (64.7 %) of long-chain (C21:0 + C23:0 + C25:0) homologues, which are claimed to prevent cardiovascular diseases and cancer. In addition, line OK72 was unique in having a C17/C21 homologue ratio as high as 0.34, likely inherited from Th. ponticum. This line along with line 280B also showed a high content of total dietary fiber. Finally, peculiar storage protein composition and kernel texture were observed in some perennial durum wheat derivatives obtained from crosses between T. turgidum subsp. durum and Th. junceiforme. This wheatgrass species was found to contain the 10-mer QQPQDAVQPF peptide, which is able to prevent prolamins from triggering inflammatory responses in celiac patients.     

Farmers perception of pesticide use and genetic erosion of landraces of tetraploid wheat (<Emphasis Type="Italic">Triticum</Emphasis> spp.) in Ethiopia

Perception of farmers’ about the use of pesticides and genetic erosion of tetraploid wheat landraces of Ethiopia was assessed through focus group discussions with farmers, on-farm observations, personal interviews with farmers, by using structured questionnaires of temporal and spatial methods. A total of 1496 farmers from seven provinces in the country were interviewed. Farmers’ knowledge about pesticide increases suggests that they are not happy on using chemicals because of their negative impact on farm land. About 75 % of the farmers believe that, although the use of pesticides may increase the production of wheat, it has a negative impact on (human) health and environment. Women showed a higher concern for pesticides’ harmfulness than men. Farmers’ valuation of genetic erosion was estimated as reduced importance of landraces, as shown by a the lower proportion of landraces either grown or sold on the market. The four most important factors cited for loss of landraces were reduction in land area per capita, displacement by released/modern varieties of hexaploid wheat and teff, reduced benefit from landraces, and displacement by other crops and chat. Genetic erosion of 100 % was observed for Triticum dicoccon in the provinces of Gojam and Gonder and for T. polonicum in Tigray and Gojam. Overall, genetic erosion in the country was 32.0, 35.3, 55.9, 84.4 and 84.4 % for T. durum Desf., T. turgidum L., T. aethiopicum Jakubz., T. polonicum L. and T. dicoccon Schrank, respectively.     

Diverse morphological and cytogenetic variation and differentiation of the two subspecies in <Emphasis Type="Italic">Aegilops geniculata</Emphasis> Roth,a wild relative of wheat

Aegilops geniculata Roth, a wild relative of wheat (2n = 4x = 28, genome UUMM), is distributed over the Mediterranean basin and nearby areas. The species consists of two subspecies, subsp. geniculata and subsp. gibberosa (Zhuk.) Hammer. The former is distributed over the whole species area and has been genetically analyzed, and the latter is endemic to Spain and North Africa and has not been genetically evaluated. In this study, to clarify the genetic variation and delineation of the two subspecies from a biosystematic viewpoint, morphological variation among 23 accessions of subsp. geniculata and three of subsp. gibberosa and chromosome pairing at meiosis and fertility in their intra- and inter-subspecific F₁ hybrids were examined. A principal component analysis based on the 11 spike characteristics clearly divided the 26 accessions into two groups representing the two subspecies. The inter-subspecific F₁ hybrids showed significantly lower frequencies of chromosome pairing, significantly higher frequencies of multivalents, and significantly lower fertilities relative to those of the intra-subspecific F₁ hybrids. It was concluded that wide-ranging cytogenetic variation is included in subsp. geniculata, that subsp. gibberosa, the intra-subspecific variation of which is small, is morphologically and cytogenetically differentiated from subsp. geniculata beyond the range of the intra-subspecific variation of subsp. geniculata, and that the two subspecies are effectively isolated reproductively by hybrid sterility. The results strongly suggested that western North Africa is one of the important diversity centers of Ae. geniculata, where two subspecies were differentiated in the past and grow together in the present.     

Characterization of a world collection of <Emphasis Type="Italic">Agropyron cristatum</Emphasis> accessions

The genetic diversity was studied of 115 Agropyron cristatum accessions from 17 countries. Tetraploids were the most common (74.8%), followed by diploid (16.3%) and hexaploid (6.9%). We observed a relation between geographic distribution and ploidy level. The tetraploids, the most widespread, were found from Europe through Russia to East Asia. The diploids appeared over the same general range, except in Turkey, Iran and Georgia where no diploid accessions were found. Hexaploid accessions mainly came from a region comprising the east of Turkey, the north of Iran and Georgia. A selection of 71 accessions, including all three ploidy levels, were analyzed by capillary electrophoresis using six wheat simple sequence repeat (SSR) markers. All markers presented high levels of polymorphism, generating 166 different alleles ranging in size between 84 and 256 bp. Based on polymorphic information content values obtained (0.579–0.968), all the SSRs were classified as informative markers (values?>?0.5). According to the dendrogram generated, all the A. cristatum accessions were distinctly classified. Diploid, tetraploid and hexaploid accessions are not clearly differentiated from each other on the basis of SSR markers. A field experiment was conducted to morphologically characterize 18 accessions including the three ploidy levels. Significant differences were found between the accessions in spike length, spike width and number of spikelets per spike. All the cytological, molecular, and morphological data demonstrate the high genetic diversity present in A. cristatum, making it a valuable resource for future breeding programs.     

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.