SSR markers and seed quality traits revealed genetic diversity in durum wheat (Triticum durum Desf.)

Molecular Biology Reports - Genetic diversity and differences among durum-wheat cultivars evolved in various regions of the world are important for sustainable production in the current climate...     

Assessment of genetic diversity among Syrian durum (Triticum turgidum ssp. durum) and bread wheat (Triticum aestivum L.) using SSR markers

Genetic diversity among 49 wheat varieties (37 durum and 12 bread wheat) was assayed using 32 microsatellites representing 34 loci covering almost the whole wheat genome. The polymorphic information content (PIC) across the tested loci ranged from 0 to 0.88 with average values of 0.57 and 0.65 for durum and bread wheat respectively. B genome had the highest mean number of alleles (10.91) followed by A genome (8.3) whereas D genome had the lowest number (4.73). The correlation between PIC and allele number was significant in all genome groups accounting for 0.87, 074 and 0.84 for A, B and D genomes respectively, and over all genomes, the correlation was higher in tetraploid (0.8) than in hexaploid wheat varieties (0.5). The cluster analysis discriminated all varieties and clearly divided the two ploidy levels into two separate clusters that reflect the differences in genetic diversity within each cluster. This study demonstrates that microsatellites markers have unique advantages compared to other molecular and biochemical fingerprinting techniques in revealing the genetic diversity in Syrian wheat varieties that is crucial for wheat improvement.     

AFLP and pedigree-based genetic diversity estimates in modern cultivars of durum wheat [ Triticum turgidum L. subsp. durum (Desf.) Husn.

A substantial amount of between and within cultivar genetic variation was detected in all the 13 registered modern Canadian durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husn.) cultivars based upon amplified restriction fragment polymorphism (AFLP). Of the approximately 950 detected AFLP markers, only 89 were polymorphic, with 41 between cultivars whereas the remaining 48 showed polymorphism within at least one cultivar. The ancestry of Canadian durum wheat cultivars was traced back to 125 cultivars, selections, and breeding lines including 17 landraces. Mean pair-wise genetic distance based on the kinship coefficient was 0.76. On the other hand, AFLP-based mean pair-wise genetic distance was 0.40. Even though there was a large difference between the means of the two diversity measures, a moderate positive correlation (r=0.457, p<0.002) was detected between the two distance matrices. Cluster analysis with the entire AFLP data divided all cultivars into three major groups reflecting their breeding origins. One group contained ’Pelissier’ alone, which was a selection from a landrace introduced into the US from Algeria. On the other hand such groupings among cultivars were not evident when KIN was used for genetic diversity measures instead. The level of genetic variation among individuals within a cultivar at the breeders’ seed level was estimated based on an inter-haplotypic distance matrix derived from the AFLP data. We found that the level of genetic variation within the most-developed cultivars is fairly substantial despite rigorous selection pressure aimed at cultivar purity in breeding programs. Comparison of AFLP and pedigree-based genetic diversity estimates in crop species such as durum wheat can provide important information for plant improvement. Received: 26 January 2001 / Accepted: 31 May 2001     

Analysis of genetic diversity of spring durum wheat (Triticum durum Desf.) cultivars released in Russia in 1929-2004

Based on genealogical analysis, the genetic diversity of 78 spring durum wheat cultivars released in Russia in 1929-2004 have been examined. The temporal trends of change in diversity were studied using series of n x m matrices (where n is the number of the cultivars and m is the number of original ancestors) and calculating coefficients of parentage in sets of cultivars released in particular years. The pool of original ancestors of spring durum wheat cultivars includes 90 landraces and old varieties, more than a half (57%) of which originate from European countries, including Russia and Ukraine (45%). The original ancestors strongly differ in the frequency of presence in the cultivar pedigrees. Landraces Beloturka, Sivouska, Kubanka (T. durum Desf.), Transbaikalian emmer, Yaroslav emmer (T. dicoccum Schuebl.), Poltavka (T. aestivum L.), and the original ancestors of cultivars Kharkov 46, Narodnaya, and Melanopus 1932 enter in the pedigrees of more than half of cultivars created within the framework of various breeding programs. At that, their distribution by cultivars from different breeding centers strongly varies. Analysis of temporal dynamics of genetic diversity, based on genetic profiles and coefficients of parentage, has shown that the genetic diversity of Russian durum wheats increased during the period examined. Nevertheless, genetic erosion of the local material-a loss of approximately 20% of the pool of Russian original ancestors-has been found. The contribution of the original ancestors to the pedigrees of different cultivars, constructed in different breeding centers and recommended for cultivation in different regions, has been estimated. The variation of the released cultivars was highest in the Lower Volga region and lowest in the Ural region. In all, the lower threshold of genetic diversity in all regions does not reach the critical level, corresponding to the similarity of half-sibs. The set of modern cultivars included in the Russian Official List 2004 has a cluster structure.     

High molecular weight glutenin subunit in durum wheat (T. durum)

Summary The diversity of high molecular weight (HMW) glutenin subunits of 502 varieties of durum wheat (Triticum durum) from 23 countries was studied using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Twenty-nine types of patterns were observed with 18 mobility bands. A total of 18 alleles were identified by comparing the mobilities of their subunits to those previously found in hexaploid wheat (T. aestivum) and in Triticum turgidum var. dicoccum. Five new alleles were detected: two on the Glu A1 and three on the Glu B1 locus. Comparison of the frequency of alleles in the three species T. aestivum, T. dicoccum and T. durum was investigated. Significant differences exist between each of these species on the basis of the frequency distributions of their three and four common alleles at the Glu A1 and Glu B1 locus, respectively. The Glu B1c allele occuring very frequently in hexaploid wheats was not found in the two tetraploid species. More than 83% of the T. durum analysed were found to have the Glu A1c (null) allele.     

Global diversity of durum wheat Triticum durum Desf. for alleles of gliadin-coding loci

Genetic diversity for the alleles of gliadin-coding loci was studied with 465 durum wheat accessions from 42 countries. A total of 108 alleles were identified for four loci; 60 alleles were described for the first time. Broad diversity of rare gliadin-coding alleles was observed. The highest genetic diversity was characteristic of durum wheat accessions from the Middle East, Trans-Caucasia, the Pyrenean Peninsula, and the Balkans. Two genetically isolated ancient branches of durum wheat were isolated. A “southern” branch included mostly accessions from the Mediterranean region, the Middle East, and Trans-Caucasia. A “northern” branch included Russian and Ukrainian durum wheat accessions and varieties obtained on their basis. An additional group included durum wheat accessions that had been obtained in several past decades on the basis of the material of international breeding centers (CIMMYT and ICARDA) and had low genetic diversity.     

A genetic linkage map of durum wheat

 A genetic linkage map of tetraploid wheat [Triticum turgidum (L.) Thell.] was constructed using segregation data from a population of 65 recombinant inbred lines (RILs) derived from a cross between the durum wheat cultivar Messapia and accession MG4343 of T. turgidum (L.) Thell. ssp dicoccoides (Korn.) Thell. A total of 259 loci were analysed, including 244 restriction fragment length polymorphisms (RFLPs), one PCR (polymerase chain reaction) marker (a sequence coding for a LMW (low-molecular-weight) glutenin subunit gene located at the Glu-B3 locus), seven biochemical (six seed-storage protein loci and one isozyme locus) and seven morphological markers. A total of 213 loci were mapped at a LOD≥3 on all 14 chromosomes of the A and B genomes. The total length of the map is 1352 cM and the average distance between adjacent markers is 6.3 cM. Forty six loci could not be mapped at a LOD≥3. A fraction (18.6%) of the markers deviated significantly from the expected Mendelian ratios; clusters of loci showing distorted segregation were found on chromosomes 1B, 3AL, 4AL, 6AL and 7AL. The durum wheat map was compared with the published maps of bread wheat using several common RFLP markers and general features are discussed. The markers detected the known structural rearrangements involving chromosomes 4A, 5A and 7B as well as the translocation between 2B-6B, but not the deletion on 2BS. This map provides a useful tool for analysing and breeding economically important quantitative traits and for marker-assisted selection, as well as for studies of genome organisation in small grain cereal species. Received: 5 January 1998 / Accepted: 31 March 1998     

Plant regeneration from protoplasts of durum wheat (Triticum durum Desf. cv. D6962)

Suspension cultures of durum wheat were established from embryogenic callus maintained in liquid medium for 30 months. Protoplasts were readily isolated from the suspension cultures with yields as high as 3 X 10⁷ protoplasts per g fresh weight suspension cells. When incubated in a modified MS medium containing half strength of the macroelements, 5 M 2,4-D (2,4-dichlorophenoxyacetic acid) and 0.6 M glucose, protoplast-derived cells divided at frequencies ranging from 1.4 to 10.0 %. After transfer to a solid subculture medium, the protoplast-derived colonies formed embryogenic protuberances, from which green plants have been regenerated.     

Genetic diversity of durum wheat as determined by AFLP in fluorescence

The DNA of fifteen Italian cultivars of durum wheat (Triticum turgidum L. ssp. durum) were analyzed by in fluorescence amplified fragment length polymorphism (fAFLP) in order to obtain the characteristic fingerprintings of genotypes and assess their genetic relatedness. Among 64 combinations of fluorescence labelled primers, three different combinations were chosen as producing a total of 6630 AFLP fragments, 2277 (34.3 %) of them being polymorphic. By using this fAFLP methodology a DNA fingerprinting of each durum wheat cultivar was generated for genotype identification. Analysis of the genetic relationships show the low variability among durum wheat cultivars.     

Biofortification of durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husnot) grains with nutrients

Durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husnot) was grown under conditions to promote mineral biofortification at the grain level. Along plant development, biomass accumulation and the kinetics of nutrients accumulation were assessed, identifying the nutrient fluxes of roots and shoots, and the timescale constraints of crop biofortification. Plants were grown under environmentally controlled conditions, submitted to four increasing concentrations of nutrient solutions (1-, 2-, 4- and 6-fold) of micro- (Fe, Zn, Cu and Mn) and macronutrients (Ca, K, P and Mg). The threshold of mineral toxicity was not reached as evaluated through plant biomass accumulation, but considering grain yield, the twofold nutrient concentration was the best treatment for biofortification. In the different treatments, the contents and the mineral unrests of roots uptake and shoots translocation varied, at different magnitudes and trends, before the onset of booting and from the physiological maturity onwards. Except for Cu, all mineral nutrients were mainly detected in the bran and embryo of the grains; therefore, the production of biofortified pasta for human consumption requires the use of integral semolina.     

Stay-green trait-antioxidant status interrelationship in durum wheat (Triticum durum) flag leaf during post-flowering

Three independent durum wheat mutant lines that show delayed leaf senescence or stay-green (SG) phenotype, SG196, SG310 and SG504, were compared to the parental genotype, cv. Trinakria, with respect to the photosynthetic parameters and the cellular redox state of the flag leaf in the period from flowering to senescence. The SG mutants maintained their chlorophyll content and net photosynthetic rate for longer than Trinakria, thus revealing a functional SG phenotype. They also showed a better redox state as demonstrated by: (1) a lower rate of superoxide anion production due to generally higher activity of the antioxidant enzymes superoxide dismutase and catalase in all of the SG mutants and also of the total peroxidase in SG196; (2) a higher thiol content that can be ascribed to a higher activity of the NADPH-providing enzyme glucose-6-phosphate dehydrogenase in all of the SG mutants and also of the NADP⁺-dependent malic enzyme in SG196; (3) a lower pro-oxidant activity of lipoxygenase that characterises SG196 and SG504 mutants close to leaf senescence. Overall, these results show a general relationship in durum wheat between the SG phenotype and a better redox state. This relationship differs across the different SG mutants, probably as a consequence of the different set of altered genes underlying the SG trait in these independent mutant lines.     

Quantitative genetic analysis and mapping of leaf angle in durum wheat

The leaf erectness profile has been used to optimize plant architecture since erect leaves can enhance photosynthesis and dry matter production by greater sunlight capture. Brassinosteroid is a recent class of phytohormones that has been related to a more erect profile. There are no reports in the literature of the genetic variability of leaf angle in doubled haploid durum wheat populations; most studies on leaf angle have focused on the inheritance. Our aim was to study the genetic variation in flag and penultimate leaf angle in a durum wheat doubled haploid mapping population, identifying and mapping quantitative trait loci influencing leaf angle. An F₁-derived doubled haploid population of 89 lines from the cross Strongfield/Blackbird was used to construct a genetic map using 423 molecular marker loci. Two greenhouse experiments and one field test were conducted using an alpha lattice in a randomized complete block design with three replicates. The leaf angle was measured on flag and penultimate leaf with a protractor at three different growth stages. The results indicated poor to moderate correlations between the position of the leaf angle and the growth stage. Transgressive segregation beyond Strongfield and Blackbird of leaf angle was observed for all environments. Putative trait loci were identified on chromosomes 2A, 2B, 3A, 3B, 4B, 5B and 7A. This work helps to understand the genetics of leaf angle in durum wheat.     

EST-SSR DNA polymorphism in durum wheat (Triticum durum L.) collections

SSRs derived from EST were molecular markers belonging to the transcribed region of the genome. Therefore, any polymorphism detected using EST-SSRs might reflect the better relationship among species or varieties. Using wheat EST-SSR markers, 60 durum wheat (Triticum durum L.) accessions from seven countries were investigated. Twenty-five primer pairs could amplify successfully in the 60 durum wheat accessions, of which tri-nucleotide repeats were the dominant type, and revealed 26 loci on all seven wheat homologous chromosome groups. A total of 87 eSSR alleles were detected, and the number of alleles detected by a single pair of primers ranged from 1 to 11, with an average of 3.3 alleles per locus. Higher numbers of alleles and PIC were identified on the B genome than those on the A genome.     

Radiation induced asynaptic mutations in durum wheat (Triticum durum DESF.)

The cytological behaviour and the inheritance of two asynaptic mutations independently obtained by neutron treatment of durum wheat seeds of var. Cappelli and Aziziah are considered. Mutant individuals of both lines have normal morphology, but are completely sterile. Analysis of chromosome behaviour at the several stages of PMC meiosis has been thoroughly accomplished in mutant progenies of both varieties during two subsequent generations. A completely random chromosome disjunction was found at anaphase I, together with multipolar spindles, laggard chromosomes etc. At meiotic second division a large amount of abnormalities were found, as a consequence of previous asynapsis and irregular polarization. Some differences in cytological behaviour were found between the asynaptic mutants of Cappelli and the asynaptics of Aziziah. - Genetic analysis demonstrated that both mutations behave as monogenic recessives.Contribution No. 139 from the Laboratory for the Application of nuclear energy to Agriculture, Casaccia Nuclear Studies Centre, Roma, Italy. - Part of the investigation was supported by Consiglio Nazionale delle Ricerche (Gruppo di lavoro per il miglioramento genetico delle piante erbacee e cultivate, Sottogruppo per il grano duro).     

A cytoplasmic male sterility (CMS) system in durum wheat

Cytoplasmic male sterility (CMS) systems are based in the incompatible interaction between nucleus and cytoplasm and are commonly used for hybrid seed production in many crop species. The msH1 CMS system in common wheat results from the incompatibility between the nuclear genome of wheat and the cytoplasm of Hordeum chilense. Fertility restoration of the CMS phenotype is associated with the addition of the short arm of chromosome 6H^ch from H. chilense. In this work, we attempt to transfer the msH1 system to durum wheat and to evaluate its potential as a new source of CMS for the production of hybrid durum wheat. For that purpose, an alloplasmic durum wheat line was developed by substituting wheat cytoplasm by that from H. chilense. This line was completely male sterile. Also, the double translocation T6H^chS·6DL was transferred from common wheat into durum wheat, to test its potential as a restorer line. Finally, the system was tested by using the double T6H^chS·6DL translocation in durum wheat as pollen donor for the alloplasmic male sterile line, which confirmed the fertility restoration ability of 6H^chS in durum wheat.     

Molecular linkage map for an intraspecific recombinant inbred population of durum wheat (Triticum turgidum L. var. durum)

Durum wheat (Triticum turgidum L. var. durum) is an economically and nutritionally important cereal crop in the Mediterranean region. To further our understanding of durum genome organization we constructed a durum linkage map using restriction fragment length polymorphisms (RFLPs), simple sequence repeats (SSRs) known as Gatersleben wheat microsatellites (GWMs), amplified fragment length polymorphisms (AFLPs), and seed storage proteins (SSPs: gliadins and glutenins). A population of 110 F₉ recombinant inbred lines (RILs) was derived from an intraspecific cross between two durum cultivars, Jennah Khetifa and Cham 1. The two parents exhibit contrasting traits for resistance to biotic and abiotic stresses and for grain quality. In total, 306 markers have been placed on the linkage map – 138 RFLPs, 26 SSRs, 134 AFLPs, five SSPs, and three known genes (one pyruvate decarboxylase and two lipoxygenases). The map is 3598 cM long, with an average distance between markers of 11.8 cM, and 12.1% of the markers deviated significantly from the expected Mendelian ratio 1:1. The molecular markers were evenly distributed between the A and B genomes. The chromosome with the most markers is 1B (41 markers), followed by 3B and 7B, with 25 markers each. The chromosomes with the fewest markers are 2A (11 markers), 5A (12 markers), and 4B (15 markers). In general, there is a good agreement between the map obtained and the Triticeae linkage consensus maps. This intraspecific map provides a useful tool for marker-assisted selection and map-based breeding for resistance to biotic and abiotic stresses and for improvement of grain quality. Received: 14 February 2000 / Accepted: 28 April 2000     

Genetic diversity of modern Russian durum wheat cultivars at the gliadin-coding loci

The allelic diversity at four gliadin-coding loci was studied in modern cultivars of the spring and winter durum wheat Triticum durum Desf. Comparative analysis of the allelic diversity showed that the gene pools of these two types of durum wheat, having different life styles, were considerably different. For the modern spring durum wheat cultivars, a certain reduction of the genetic diversity was observed compared to the cultivars bred in the 20th century.     

Catalogue of alleles of gliadin-coding loci in durum wheat (Triticum durum Desf.)

Gliadins are seed storage proteins which are characterized by high intervarietal polymorphism and can be used as genetic markers. As a result of our work, a considerably extended catalogue of allelic variants of gliadin component blocks was compiled for durum wheat; 74 allelic variants for four gliadin-coding loci were identified for the first time. The extended catalogue includes a total of 131 allelic variants: 16 for locus Gli-A1(d), 19 for locus Gli-B1(d), 41 for locus Gli-A2(d), and 55 for locus Gli-B2(d). The electrophoretic pattern of the standard cultivar and a diagram are provided for every block identified. The number of alleles per family is quite small for loci Gli-A1(d) and Gli-B1(d) of durum wheat, as contrasted to loci Gli-A2(d) and Gli-B2(d) that are characterized by large families including many alleles. The presence of large block families determines a higher diversity of durum wheat for loci Gli-A2(d) and Gli-B2(d) as compared to Gli-A1(d) and Gli-B1(d). The catalogue of allelic variants of gliadin component blocks can be used by seed farmers to identify durum wheat cultivars and evaluate their purity; by breeders, to obtain homogenous cultivars and control the initial stages of selection; by gene bank experts, to preserve native varieties and the original biotypic composition of cultivars.