Characterization of tetraploid wheat germplasm for resistance to Pseudocercosporella herpotrichoides, cause of eyespot disease

Eyespot disease, caused by Pseudocercosporella herpotrichoides, can be devastating to winter wheat grown in northern Europe and the northwest USA. Accessions from eight different tetraploid wheat species randomly extracted from core collections were scored for resistance to eyespot disease using a -glucuronidase (GUS)-transformed strain of P. herpotrichoides. The GUS values for the combined population followed a quasi-Gaussian distribution. Three species, Triticum dicoccoides, T. durum and T. turanicum, showed significant variation (P < 0.001) in disease response with T. dicoccoides having the lowest disease scores, i.e. highest levels of resistance. All tetraploid accessions were less resistant than resistant diploid T. tauschii accessions. Thirteen percent of tetraploid accessions had disease scores that ranged between the average of the resistant accessions of T. tauschii and the moderately resistant hexaploid germplasm line Cerco. Eight accessions (three accessions of T. dicoccoides, two of T. turgidum and three of T. durum) with low disease scores (resistance) to infection were selected for further genetic analysis.     

Variation in B-LMW glutenin subunits in Einkorn wheats

A collection of 50 Einkorn wheat accessions (26 of T. monococcum, 14 of T. baeoticum and 10 T. urartu) was analyzed for low-molecular-weight (LMW) glutenin subunit composition. At the B-LMW glutenin region, T. monococcum possessed 1 or 2 subunits, and T. baeoticum and T. urartu from 1 to 3 subunits. Seven different electrophoretical patterns were detected among T. monococcum and six in both T. baeoticum and T. urartu. T. monococcum was shown to be the less polymorphic, followed by T. baeoticum and by T. urartu. The fact that more glutenin genes are expressed in Einkorn wheats than in polyploid cultivated wheats suggests their possible use in breeding for improving bread and durum wheat quality.     

New descriptions of hulled wheats on the infraspecific level

New morphological variation was detected in the hulled wheats Triticum monococcum L. (cultivated), T. urartu Thum. ex Gandil. (wild) and T. baeoticum Boiss. em. Schiemann (wild) and four, five and six new botanical varieties were described, respectively. This formal approach complements the classical infraspecific classifications.     

Resistance to wheat leaf rust in Aegilops tauschii Coss. and inheritance of resistance in hexaploid wheat

A collection of 164 Aegilops tauschii accessions, obtained from Gatersleben, Germany, was screened for reaction to leaf rust under controlled greenhouse conditions. We have also evaluated a selection of synthetic hexaploid wheats, produced by hybridizing Ae. tauschii with tetraploid durum wheats, as well as the first and second generation of hybrids between some of these resistant synthetic hexaploid wheats and susceptible Triticum aestivum cultivars. Eighteen (11%) accessions of Ae. tauschii were resistant to leaf rust among which 1 was immune, 13 were highly resistant and 4 were moderately resistant. Six of the synthetic hexaploid wheats expressed a high level of leaf rust resistance while four exhibited either a reduced or complete susceptibility compared to their corresponding diploid parent. This suppression of resistance at the hexaploid level suggests the presence of suppressor genes in the A and/or B genomes of the T. turgidum parent. Inheritance of leaf rust resistance from the intercrosses with susceptible bread wheats revealed that resistance was dominant over susceptibility. Leaf rust resistance from the three synthetics (syn 101, syn 701 and syn 901) was effectively transmitted as a single dominant gene and one synthetic (syn 301) possessed two different dominant genes for resistance.     

Molecular characterization of a novel waxy allele (Wx-A u 1a) from Triticum urartu Thum. ex Gandil.

Granule Bound Starch Synthase I, or waxy protein, is the sole enzyme responsible for the accumulation of amylose during the development of starch granules in wheat. The full coding region of the waxy (Wx) gene was sequenced in Triticum urartu, (a wild diploid species) and is related to the A genome of polyploid wheats. The Wx gene of T. urartu (Wx-A ^u 1) showed a homology of ~88.0?% with Wx-A1 from polyploid wheats. A greater homology was found with Wx-A ^m 1 from the diploid cultivated wheat einkorn. Most of the differences were found in introns although several changes were also detected in exons that led to amino acid changes in the transit peptide and mature protein. These results show the potential of T. urartu as a source of new alleles that could be used in the breeding of durum and common wheat in order to synthesize starches with different properties.     

Polymorphism of high M r glutenin subunits in wild emmer Triticum turgidum subsp. dicoccoides: chromatographic,electrophoretic separations and PCR analysis of their encoding genes

Triticum turgidum subsp. dicoccoides (Körn. ex Asch. et Graebn.) Thell. (AABB), the immediate progenitor of tetraploid and hexaploid wheats, is a species characterised by a wide range of protein polymorphism and by high protein content. Surveys on polymorphism and genetic control of the high molecular weight glutenin subunits (HMW-GS) present in this species, in two forms x- and y-type at the Glu-A1 and Glu-B1 loci, are still considered useful, both to improve technological properties of breeding varieties and to study the genome evolutionary process in wheats. Comparative Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoretic and Reversed Phase High Performance Liquid Chromatographic analyses (SDS-PAGE, RP-HPLC) of the HMW-GS present in several accessions of T. turgidum subsp. dicoccoides allowed the detection of new alleles of Glu-A1 and Glu-B1 loci, with x- and y-type glutenin subunits, apparently similar to those present in cultivated wheats in molecular weight, but different in surface hydrophobicity. In addition, changes in the number of x- and y-type subunits at the glutenin loci were also ascertained. The y-type subunits at the Glu-A1 locus, which are never expressed in cultivated bread and durum wheats, and single y-type expressed glutenin subunits at the Glu-B1 locus were also identified in several accessions. DNA extracted from samples, differing in number or type of HMW-GS and corresponding to x- and y-type genes at Glu-1 loci, were amplified using specific primers, two of which were constructed within the transposon-like sequence of Chinese Spring DNA and analysed by polymerase chain reaction. The results showed this insertion in some accessions of T. turgidum subsp. dicoccoides and also the presence of silent Ax, Bx and By type genes. The usefulness for breeding of these comparative analyses carried out on different HMW-GS alleles detected in Triticum turgidum subsp. dicoccoides, is discussed.     

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.     

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.     

Analysis of fatty acid steryl esters in tetraploid and hexaploid wheats: identification and comparison between chromatographic methods

Fatty acid steryl esters (FASE) in whole meal of 14 genotypes of tetraploid wheats (Triticum dicocconand T. durum) and 17 genotypes of hexaploid wheats (T. spelta and T. aestivum) were analyzed using different chromatographic strategies. By both GC-FID and HPLC-ELSD, tetraploid wheats are lacking two major peaks. The amounts of FASE, calculated on the basis of the GC-FID analysis, were double in hexaploid species as compared to tetraploids (40 and 20 mg/100 g db, respectively). HPLC with ESI-MS detection enabled the identification of FASE by the characteristic fragmentations and ion-adducts of each molecule. The distribution of steryl residues was not different between the wheat species: the main class of steryl derivatives found was the beta-sitosteryl derivatives, followed by campesteryl derivatives with small amounts of stigmasteryl esters. The esterified fatty acids explain the difference between the hexaploid and tetraploid wheats. In particular, small amounts of campesteryl and beta-sitosteryl, while no trace of stigmasteryl palmitates, were found in T. durum or its hulled ancestor T. dicoccon. Steryl oleates were not detectable in T. aestivum or its hulled ancestor T. spelta, which is consistent with the filogenesis of tetraploid and hexaploid species. Both chromatographic techniques (GC and HPLC) showed that FASE are useful to discriminate between hexaploid and tetraploid wheats from both qualitative and quantitative points of view.     

Evaluation of emmer wheat and other Triticeae for resistance to Russian wheat aphid

Summary During the winters of 1990/91 and 1991/92, 181 accessions of Triticum dicoccon Schrank from the CIMMYT gene-bank were screened in the field for resistance to Russian wheat aphid, Diuraphis noxia (Kurdjumov). Accessions were sown in hill plots of 10 seeds and artificially infested with D. noxia at the two-leaf growth stage. Hills were visually assessed for damage at tillering, booting and heading. Entries differed significantly in their reaction to D. noxia, and severity of symptoms increased with time. Twenty four of the entries were highly resistant to the aphid. In winter 1991/92, 807 accessions of wild and cultivated wheats (26 species) and synthetic hexaploids were screened similarly for resistance to D. noxia. A large number of A-genome species were resistant, while few D-genome species were identified as resistant. These newly discovered sources of resistance can be used to expand the genetic base of resistance to D. noxia in both bread (T. aestivum L.) and durum wheats (T. turgidum L. convar. durum (Desf.) Mackey).     

Biodiversity of Diploid D-Genome Aegilops Tauschii Coss. in Iran Measured Using Microsatellites

Microsatellite markers were used to analyse the biodiversity of 57 accessions of different subspecies and varieties of wild Aegilops tauschii (2n = 2x = 14; D genome) collected across the major areas where it grows in Iran. Levels of diversity were high, with numbers of alleles averaging 7.3 (ranging up to 12) and polymorphism information contents averaging 0.6591. One accession was notably more similar to two of the D genome in hexaploid wheats (Triticum aestivum) used as outgroups. Within the Ae. tauschii accessions, no markers were characteristic for taxa or geographical origin, suggesting high gene flow between the subspecies and varieties, although some groupings, which could be related to geographical origin, were evident. This survey demonstrates the high diversity present in wild goatgrass in Iran, and indicates that there is value in sampling for useful genes for wheat breeding.     

Identification of genomic constitutions of <Emphasis Type="Italic">Oryza</Emphasis> species with the B and C genomes by the PCR-RFLP method

Oryza officinalis complex is the largest and the most complicated group in the genus Oryza L., consisting of about ten species with the B, C, BC, CD, and E genomes. Taxonomy and identification of the species, particularly those with the B, C and BC genomes, are difficult due to the similar morphology and overlapping distribution of some species. The difference in ploidy levels of some species adds more complexity. In the present study, we surveyed 64 accessions of rice germplasm in the O. officinalis complex using RFLP analysis of PCR-amplified Adh genes in addition to chromosome counting. The results confirmed that all O. rhizomatis accessions are diploids with the C genome, whereas all O. minuta accessions are tetraploids having the BC genome. However, both diploid and tetraploid forms were found for the accessions identified in the genebank as O. officinalis, O. punctata and O. eichingeri. The tetraploid form of O. officinalis with the BC genome is exclusively distributed in India and has been described as O. malampuzhaensis. The tetraploid form of O. punctata which has been called O. schweinfurthiana by some workers was found to be as widely distributed as its diploid form in Africa. It is noteworthy that two accessions that had been determined as tetraploid O. officinalis were actually belonging to a species with the CD genome (O. latifolia). Our results promote a better understanding of the genomic constitutions of many accessions in the O. officinalis complex and correct determination of the genebank material, which serves as an important basis of germplasm cataloguing for further research and utilization.     

Microsatellite markers – a new tool for distinguishing diploid wheat species

Triticum baeoticum and T. urartu are very similar morphologically. By using microsatellite markers it was possible to distinguish between these two species. Microsatellite markers are, therefore, a powerful new tool to support the determination of critical races in diploid wild wheat species. They also allow the discussion of evolutionary pathways within Triticum.     

Grain Quality of Emmer Wheat Derived Synthetic Hexaploid Wheats

The wild diploid goat grass (Aegilops tauschii Cosson), and the cultivated tetraploid emmer wheat (Triticum turgidum L. subsp. dicoccon (Schrank) Thell.) may be important sources of genetic diversity for improving hexaploid bread wheat (Triticum aestivum L.). Through interspecific hybridization of emmer wheat and Ae. tauschii, followed by chromosome doubling, it is possible to produce homozygous synthetic hexaploid wheat. Fifty-eight such synthetic hexaploids were evaluated for grain quality parameters: grain weight, length, and plumpness, grain hardness, total protein content, and protein quality (SDS-Sedimentation volume, SDS-S). Most synthetics showed semi-hard to hard grain texture. Results showed significant genetic variation among the synthetic hexaploids for protein content, SDS-S values, and grain weight and plumpness. Quality measurement values of synthetic hexaploids were regressed on corresponding values of the emmer wheat parents. With this offspring-parent regression, protein content and SDS-S values explained 8.7 and 28.8%, respectively, of the variation among synthetics, indicating a significant contribution from the emmer wheat parents for these traits. The synthetic hexaploids, in general, had significantly higher protein content (15.5%, on average) and longer grains than ‘Seri M82’, the bread wheat control (13.1% protein content). Synthetics with SDS-S values and grain weights higher than those of ‘Seri M82’ were also identified. Protein content among synthetics showed significantly negative correlations with grain weight and plumpness, but no correlation with SDS-S values. Despite these negative correlations, 10 superior synthetic hexaploid wheats, derived from nine different emmer wheat parents and with above average levels of protein content, SDS-S values, and either grain weight or plumpness, were identified. This study shows that genetic variation for quality in tetraploid emmer wheat can be transferred to synthetic hexaploid wheats and combined with plump grains and high grain weight, to be used for bread wheat breeding.     

Variation in salt tolerance within a Georgian wheat germplasm collection

Bread wheat Triticum aestivum L. possesses a genetic variation for the ability to survive and reproduce under salt stress conditions. Durum wheat (T. durum Desf.) is in general more sensitive in comparison to bread wheat, however, exceptions can be found showing the same extent of salt tolerance. Endemic wheats in general are characterised by a high adaptability to their environment. The level and variability of salt tolerance were assessed in a germplasm collection of 144 winter and spring wheat accessions from Georgia comprising Triticum aestivum L., T. durum Desf., T. dicoccon Schrank, T. polonicum L. and Georgian endemics: T. carthlicum Nevski, T. karamyschevii Nevski, T. macha Dekapr. et Menabde, T. timopheevii (Zhuk.) Zhuk. and T. zhukovskyi Menabde et Ericzjan. The accessions were tested for salt tolerance at the germination stage. Large variability in salt tolerance within the Georgian germplasm was found among the different wheat species. The endemic hexaploid winter wheat T. macha and the endemic tetraploid wheat T. timopheevii were among the most tolerant materials, thus presenting promising donors for salt tolerant traits in future breeding efforts for salinity tolerance in wheat.     

DNA Fingerprinting and Genetic Characterization of Anatolian Triticum spp. using AFLP Markers

In this study, genetic analysis of Triticum spp. was carried out using AFLP markers. Six AFLP selective combinations were scored as presence and absence of bands for all the individual samples obtained from a single seed of each accession (70 accessions); T. baeoticum (21), T. monococcum (5), T. urartu (16), T. araraticum (7), T. dicoccoides (16) and T. dicoccon (5), resulting in 506 polymorphic AFLP bands. The phylogenetic tree showed two major clusters; one was composed of T. monococcum (AA) and T. baeoticum (AA), and the other cluster included T. araraticum (AAGG), T. dicoccon (AABB), T. dicoccoides (AABB), and T. urartu (AA). T. urartu, although having a diploid AA genome, did not cluster with other A genome diploids such as T. monococcum and T. baeoticum; instead it clustered together with the tetraploid species, confirming that T. urartu is the A genome progenitor. The extent of variations within and among species is discussed.     

Natural and human selection for purple-grain tetraploid wheats in the Ethiopian highlands

Summary Purple-grain tetraploid wheats (Triticum turgidum L.) are widely cultivated in the Ethiopian highlands despite the claim that they have lower industrial quality properties and market prices than the white or red/brown seed-colour types. In an attempt to find a possible explanation for this, the three seed-colour groups were compared for grain yield, other 11 agronomic traits and protein content. Five traits displayed significant differences between seed colour groups where the purple-seed was superior; earlier maturity, shorter height, and higher fertility, tillering capacity and harvest index. Most of these are important adaptive traits to waterlogging stress on dark-clay soils (pellic vertisols) where the great bulk of the Ethiopian tetraploid wheats have been grown. Furthermore, among the three seed-colour groups, purple-seed wheat has the best malting quality for the preparation of arekie, a locally distilled spirituous liquor. It, therefore, appears that both natural and human selections have been reponsible for their continued cultivation. Hence, the notion that purple-seeded wheat is the “least preferred” should be interpreted carefully not to necessarily address the whole community in Ethiopia. As to their taxonomy, all tetraploid wheat taxa (T. turgidum L. sensu lato, 2n = 4x = 28) that are found in Ethiopia, with the possible exception of T. dicoccon Shrank (locally known as Adja), may possess the purple pericarp-colour, although in varying frequencies; very low inT. polonicum L., and high inT. carthlicum Nevski andT. durum Desf.     

Wheat artificial amphiploids involving the <Emphasis Type="Italic">Triticum timopheevii</Emphasis> genome: their studies,preservation and reproduction

The effective utilisation of available genetic resources of related species is essential for successful crops breeding and maintaining genetic variability within crops. Bread wheat, the basic cultivated wheat species, is an amphiploid (2n = 6x = 42) and, therefore, the production of new synthetic amphiploids using genomes of related species should reduce the difficulties caused by direct crossings, for example, between hexaploid wheat and diploid relatives. Hence, exploiting synthetic amphiploids is an effective and rapid way of introgressing desirable traits from related species into cultivated wheats. Some of the artificial amphiploids that already exist were produced 80 years ago. Yet little work has been done to highlight potential contamination and/or genetic changes during their conservation by genebanks. Thus, we utilised the electrophoresis of wheat endosperm storage proteins (gliadins) to check such amphiploid authenticity, and also where differences had been previously observed between synthetic wheat amphiploids. In addition, we checked putative amphiploid accessions where Triticum timopheevii (GGA^tA^t) was recorded as one of the parents. A synthetic species, T. timococcum produced by Kostov, together with a natural T. zhukovskyi found in Georgia (the former Soviet Union) were revealed to be identical according to our assays. The existence of several T. kiharae accessions independently produced by different authors was confirmed, and they exhibited polymorphism for a number of traits, including spike characters (awning, hairy glumes) and growth habit (spring vs. winter). The effective conservation of artificial amphiploids in genebanks is discussed.     

野生二粒小麦导入普通小麦抗白粉病基因MlWE27的鉴定和分子标记

由白粉病菌(Blumeria graminis f.sp.tritci)引起的小麦白粉病是严重影响小麦安全生产的主要病害之一.本研究将来自以色列的野生二粒小麦(Triticum dicoccoides)WE27的坏白粉病基因通过杂交和连续回交,导入普通小麦遗传背景中,育成高抗白粉病小麦新品系3D256(其系谱为燕大1817/WE27//农大015/3/941,F6).将3D256和高感小麦白粉病的普通小麦品系薛早配制杂交组合,对其F_1、F_2分离群体和F_3 家系进行白粉病抗性鉴定和遗传分析.结果表明,3D256携带抗白粉病显性单基因,暂命名为MlWE27.利用集群分离分析法(RSA)和分子标记分析,发现3个SSR标记(Xwmc243、Xwmc 154和Xbarc318)、1个EST-SSR标记(Xdp357)、1个AFLP转化的SCAR标记(XCAUG1)和1个RFLP探针转化的STS标记(XWG516-1)与抗白粉病基因MlWE27连锁,在连锁图上的顺序为Xdp357-Mlwe27-XCAUG1-XWG516-1-Xwmc243-Xwmc154-Xbarc318.利用中国春缺体-四体系、双端体系和缺失系将抗白粉病基因MlWE27定位于染色体2B短臂的末端Bin0.84-1.00上.这一普通小麦抗白粉病种质资源的创制及其连锁分子标记的建立为小麦抗病基因分子标记辅助选择、基因积聚和分子育种提供了新的物质基础.     

Assessment of variability in salt tolerance based on seedling growth in Triticum durum Desf.

Variability in response to salinity was examined in 29 Triticum durum Desf. accessions or cultivars based upon relative root and shoot lengths of 14-day-old seedlings grown in control and 100 mM NaCl solutions. NaCl caused a significant reduction of root and shoot growth for all accessions/cultivars, but the degree of reduction differed between them. Some cultivars had significantly greater relative root and shoot lengths than others, suggesting that there may be potentially useful variability in salinity tolerance within the durum wheats.