Relationship between Quality, Colour of Glume and Gliadin Electrophoregrams in Durum Wheat

Gliadin electrophoregrams, protein content and SDS-sedimentation values from the modified SDS-sedimentation test were obtained from six Greek cultivars of Triticum durum and 98 biotypes (78 with white glumes, 20 with red glumes). Our results provide wheat breeders with the tool TO predict the cooking quality of durum wheat from early generation breeding lines, since we were able to correlate the colour of the glume with the presence or absence of gliadin bands 42/45. All red glume biotypes lacked gliadin band 45 and possessed gliadin band 42. Furthermore, their SDS sedimentation values were around 27 (weak gluten), indicating poor cooking quality. Yet, the electrophoregrams of the gliadin proteins allowed an identification of the wheat cultivars examined, since for each cultivars the pattern was different. No correlation was found between the colour of the glume and the amount of total gram protein.     

Gliadins as biochemical markers for Aegilops turcomanica genes in wheat lines

An Aegilops turcomanica-typical gliadin was discovered in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns of ethanol-soluble seed proteins from wheat lines Tr. 476, 482 and 492, which had been derived from a direct cross of Ae. turcomanica and Triticum aestivum and which revealed powdery mildew resistance due to a putative Ae. turcomanica-introgression. This Ae. turcomanica-derived gliadin was tested for its suitability as biochemical marker. For this purpose, doubled-haploid lines were produced via anther culture from crosses of Tr. 482 and Tr. 492 with actual winter wheat cultivars and breeding lines. Until now, 173 lines with Ae. turcomanica-gene(s) have been selected from 297 doubled haploid wheat lines.     

Resistance in spelt wheat to yellow rust

Summary Seven spelt wheat accessions of different origin were hybridized with the susceptible bread wheat cultivar Taichung 29 in order to study the genetics of their resistance to yellow rust (Puccinia striiformis Westend. f. sp. tritici). One Iranian and five European accessions were found to carry Yr5 of Triticum aestivum ssp. spelta var. album, whereas a factor for resistance in the Iranian accession 415 was confirmed to be genetically distinct from Yr5. The alleles for resistance in each of the accessions studied showed a monogenic dominant mode of inheritance. Twenty-eight spelt wheat accessions, including those studied for their resistance to yellow rust, were subjected to polyacrylamide-gel-electrophoresis to study variation for gliadin storage protein patterns. Thirteen distinct patterns were revealed, implying the presence of duplicates within the studied spelt wheat collection.     

Gliadin allele composition of Yugoslav winter wheat cultivars

Summary The complete gliadin allele composition of 57 Yugoslav common winter wheat cultivars was studied. Large differences were found in gliadin genotypes among cultivars bred at different Yugoslav breeding centres as well as between early and recent wheats bred in the Novi Sad breeding centre. Yugoslav wheats have limited variation in gliadin alleles, of which Gli-A1a, Gli-B1b, Gli-B1l, Gli-D1b, Gli-A2e, Gli-B2b, Gli-D2a are shown to be the most frequent. Examples of favoured alleles to new cultivars are described. 40% of the studied wheat cultivars have natural biotypes differing in gliadin allele composition. The frequency of cultivars with the biotypes differs greatly among breeding centres.     

Genetic mapping of sedimentation volume across environments using recombinant inbred lines of durum wheat

SDS-sedimentation volume (SV) is a biochemical index widely used to evaluate flour quality in durum and bread wheats. Significant association between SV and endosperm proteins (gliadin, high-molecular-weight- and low-molecular-weight-glutenin subunits) have been reported. Protein loci, however, account for only a portion of the total genetic variability. The objective of this study was to identify and locate quantitative trait loci (QTLs) associated with SV in a set of recombinant inbred (RI) lines, derived from a cross between the cv.‘Messapia’ of durum wheat and the accession MG4343 of the var. dicoccoides, and characterized for 259 genetic and molecular (RFLP) markers. Significant differences were detected for the quality index in the six environments examined, while the pattern of variability was that of a quantitative trait. Regression analysis of marker loci and sedimentation volume indicated, as expected, that chromosome 1B, on which are located the Gli-B 1/Glu-B 3 loci for some gliadin and glutenin subunits, is important for wheat quality. Two additional regions located on chromosomes 6AL and 7BS, and four regions on 1AL, 3AS, 3BL and 5AL, were shown to have single-factor effects on sedimentation volume at P < 0.001 and P < 0.01, respectively. Positive effects were contributed by both parents. A multiple linear regression model consisting of seven significant loci on different chromosomes explained 62–91% of the genotypic variation of the trait. The availability of linked markers to QTLs may facilitate the genetic dissection of quantitative traits and the early selection in wheat breeding programmes.     

Genetic polymorphisms at Gli-Dt gliadin loci in Aegilops tauschii as revealed by acid polyacrylamide gel and capillary electrophoresis

Gliadin variation at Gli‐D^t1 and Gli‐D^t2 loci in 198 Aegilops tauschii accessions was studied by acid polyacrylamide gel electrophoresis (A‐PAGE) and capillary electrophoresis (CE). High genetic polymorphisms were found at both gliadin coding loci, revealing a total of 184 and 169 gliadin variants at the Gli‐D^t1 and Gli‐D^t2 loci, respectively. In particular, 12 gliadin blocks encoded by different alleles were apparently expressed and readily identified in six synthetic hexaploids produced by hybridization between Triticum durum and Ae. tauschii accessions. Compared with Ae. tauschii ssp. eusquarrosa, the gliadin profile of the D genome in Ae. tauschii ssp. strangulata more resembles that of T. aestivum, supporting the view that the subspecies strangulata is the most likely progenitor of bread wheat. Capillary electrophoresis analysis showed that the method is capable of separating and characterizing gliadins with speed, in high resolution using small sample amounts, and is well‐suited to detect protein alleles and to identify desirable genotypes in wheat quality improvement.     

Allelic variation at the gliadin coding loci of improved Ethiopian durum wheat varieties

The objective of this study was to determine gliadin allele compositions of 20 improved Ethiopian durum wheat varieties using acid-polyacrylamide gel electrophoresis (A-PAGE). Each block of co-dominantly inherited polypeptides encoded by gliadin loci were identified and their genetic diversities were estimated using statistical analyses. A total of 30 electrophoretic blocks were identified at five major gliadin loci. In addition, four novel gliadin blocks were identified. Gli-B1 and Gli-A2 loci had higher numbers of gliadin alleles (nine and ten, respectively) compared to other loci. Alleles Gli-A1c on chromosome 1A, Gli-B1c on chromosome 1B, Gli-A2a, and Gli-A2o on chromosome 6A, and Gli-B2h on chromosome 6B had maximal frequencies in their corresponding loci. Varieties were classified into three main clusters and one singleton based on genetic distances of detected gliadin alleles. These results indicate that Ethiopian durum wheat varieties are genetically diverse with unique allele compositions at gliadin-coding loci.     

Cytogenetic analysis of a long-term Triticum timopheevii (Zhuk.) Zhuk. cell suspension culture

In vitro culturing of plant cells can cause changes in karyotype. Chromosome variations following long-term propagation in suspension culture of Triticum timopheevii (Zhuk.) Zhuk. were studied by routine staining and C-banding. The culture was highly heterogeneous with respect to the number and structure of chromosomes. The modal class cells had a lower chromosome number than T. timopheevii (2n= 28). This data was confirmed by cytophotometric analysis of nuclear-DNA content. Frequencies of chromosome loss varied for different homoeologous groups. A^t genome chromosomes tended to be preferentially eliminated in cells of different ploidy levels. Deletions, insertions, translocations, telocentric chromosomes, isochromosomes and dicentrics and their derivatives were observed in cultured cells. Chromosomes of various homoeologous groups differed in the frequencies and spectra of re—arrangements, but most aberrations occurred in the G-genome chromosomes. In vitro chromosome modifications did not correspond to in vivo variation. Presumably, this difference was caused by differences in the mechanisms of adaptation to the environment at the levels of the cell and the whole organism. G-genome chromosomes were more frequently involved in this process, both in vivo and in vitro.     

Expression of morphological and flowering time variation through allopolyploidization: an empirical study with 27 wheat synthetics and their parental Aegilops tauschii accessions

It was recently shown that allopolyploidy brings novel epistatic interactions to genes belonging to different genomes. However, systematic studies of the phenotypic relationships between synthetic hexaploid wheats and their parental lines have not been conducted. In this study, 27 synthetic hexaploid wheats were produced by crossing the tetraploid wheat cultivar ‘Langdon’ with 27 accessions of Aegilops tauschii. Variations in 20 morphological and flowering traits were analysed in both the synthetic wheat lines and the parental Ae. tauschii accessions. The 20 traits exhibited large variations in the wheat lines. For many of the traits, the degree of variation in the parental accessions was reduced in the hexaploid derivatives. Principal component analysis of floret‐related traits divided the Ae. tauschii accessions into two subspecies, ssp. tauschii and ssp. strangulata, but this parental pattern of subspecific division was not detectable in the hexaploids. Our results suggest that the ‘Langdon’ genome may have an alleviating effect on the expression of D‐genome‐derived variations in derived synthetics.     

In vitro screening and field evaluation of tissue-culture-regenerated sorghum (Sorghum bicolor (L.) Moench) for soil stress tolerance

Summary Sorghum bicolor (L.) Moench is generally quite sensitive to salt and acid (high aluminium) soil stresses, but quite tolerant of drought stress. As with any stress phenomenon, intra-specific variability exists within the genus. In vitro cell selection and somaclonal variation offer an alternative to traditional breeding methodology for generating improved breeding lines for hybrid development. A field selection protocol was developed for the three soil stresses and inter-stress evaluations were conducted in an effort to find multiple, stress-tolerant genotypes. The acid soil-drought stress, super-tolerant selections were located by the R₇ generation when exposed to a combined aluminium-drought stress field environment and when the regeneration population (number of regenerated lines from one callus source) was maintained at 15,000 plants or higher. A variant frequency of 0.1 to 0.2% for stress tolerance and acceptable agronomic traits among the surviving somaclones, provided an adequate number of phenotypes with desirable agronomic characteristics and a high level of soil stress tolerance. Subsequent research verified that the stress-tolerant regenerants had superior acid soil and drought stress tolerance to that of the donor parents, that their yield capabilities under stress were superior to their parents, and that their stress tolerance attributes were transferred in hybrid combinations. In vitro selection was not effective in increasing the number of field stress survivors. In fact, superior germplasms were developed from non-stressed callus or salt-stressed callus. In vitro selection reduced regeneration frequency and subsequent survival of plants under field stress. In vitro-stressed regenerants should be subjected only to non-stressed environments to maintain population numbers for field selection and thereafter should be subjected to stress environments during later (R₅₊) generations. The optimal strategy for the exploitation of somaclonal variation may be through short-term cell culture (< 12 months) with no attempt at in vitro selection.     

Quantitative trait locus analysis of wheat quality traits

Milling and baking quality traits in wheat (Triticum aestivum L.) were studied by QTL analysis in the ITMI population, a set of 114 recombinant inbred lines (RILs) generated from a synthetic-hexaploid (W7985) × bread-wheat (Opata 85) cross. Grain from RILs grown in U.S., French, and Mexican wheat-growing regions was assayed for kernel-texture traits, protein concentration and quality, and dough strength and mixing traits. Only kernel-texture traits showed similar genetic control in all environments, with Opata ha alleles at the hardness locus Ha on chromosome arm 5DS increasing grain hardness, alkaline water retention capacity, and flour yield. Dough strength was most strongly influenced by Opata alleles at 5DS loci near or identical to Ha. Grain protein concentration was associated not with high-molecular-weight glutenin loci but most consistently with the Gli-D2 gliadin locus on chromosome arm 6DS. In Mexican-grown material, a 2DS locus near photoperiod-sensitivity gene Ppd1 accounted for 25% of variation in protein, with the ppd1-coupled allele associated with higher (1.1%) protein concentration. Mixogram traits showed most influence from chromosomal regions containing gliadin or low-molecular-weight glutenin loci on chromosome arms 1AS, 1BS, and 6DS, with the synthetic hexaploid contributing favorable alleles.Some RI lines showed quality values consistently superior to those of the parental material, suggesting the potential of further evaluating new combinations of alleles from diploid and tetraploid relatives, especially alleles of known storage proteins, for improvement of quality traits in wheat cultivars.Contribution number 06-77J from the Kansas Agricultural Experimental Station.     

Improvement of in vitro culture response of elite wheat cultivars by selecting the source spike, the scutellum size and the temperature for the induction of embryogenesis

High frequencies of regeneration are desirable characteristics of in vitro culture from elite wheat cultivars. The effect of the source spike, the scutellum size and the induction temperature on embryogenesis, regeneration and number of plants per explant were optimized in three varieties of wheat and later determined in fifteen varieties of both bread and durum wheat. The highest frequencies of embryogenesis, regeneration and plants per explant were obtained when the first spike was used as source of scutella and decreased using the scutella from the second and third spikes. Scutella from the third spike yielded half the number of plants per explant than those from the first spike. Significant differences were also found for embryogenesis, regeneration and plants per explant as affected by scutellum size. The number of plants per explant drastically decreased in commercial varieties when scutella larger than 2.0 mm were used. The optimum induction temperature was 21 and 25°C for commercial and model genotypes, respectively. However, no differences were found for the number of plants per explant as affected by the induction temperature. The in vitro culture response of twelve and four cultivars of, respectively, bread and durum wheat were then determined. In all wheat varieties tested embryogenesis, regeneration and plants per explant varied over a wide range of values. In elite wheat varieties the selection of the source spike and the scutellum size can yield high frequencies of embryogenesis and regeneration and high numbers of plants per explant.     

Spike pretreatments, anther culture conditions, and anther culture response of 17 German varieties of spring wheat (Triticum aestivum L.)

The aim of this work was to establish an in vitro regeneration system from anther cultures of different German varieties of spring wheat (Triticum aestivum L.). Using ‘Nandu’ the most widely grown spring wheat cultivar in Germany, different culture conditions were investigated with regard to their influence on anther culture response. The best results were obtained when applying a cold pretreatment to the donor spikes and using the synthetic L3 induction medium, liquid or solidified with gelrite. The highest rates obtained in these experiments with ‘Nandu’ were 8.6% responding anthers, 22.3% embryoid induction, 15.3% albino regeneration and 5.5% green plant regeneration (all rates related to the number of cultured anthers). Of the ‘Nandu’ plants analysed, 51.1% were haploid and 44.3% were diploid, probably as a consequence of spontaneous chromosome doubling. When screening a further 16 commercial German varieties of spring wheat, 10 exhibited good anther culture response and four of these (‘Eta’‘Jondolar’, ‘Mieka’, and ‘Star’) proved to be highly responsive, reaching embryoid induction rates between 4.3 and 10.3% and rates of green plant regeneration between 5.4 and 10.7%.     

Detection of wheat-barley translocations by genomic in situ hybridization in derivatives of hybrids multiplied in vitro

Wheat-barley translocations were identified by genomicin situ hybridization (GISH) in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) × barley (Hordeum vulgare L. cv. Betzes) hybrids. The regenerated hybrids were pollinated with the wheat line Martonvásári 9 kr1. Five translocated wheat-barley chromosomes were recovered among 51 BC₂F₂ progeny from the in vitro regenerated wheat × barley hybrids. All were single breakpoint translocations with the relative positions of the breakpoints ranging from the centromere to about 0.8 of the relative arm length. Of the four translocations with intercalary breakpoints, three were transfers of terminal barley segments to wheat chromosomes; one was a transfer of a terminal wheat segment to a barley chromosome. Because of the absence of diagnostic N-bands, the identity of three barley segments could not be determined; in one translocation the barley chromosome involved had a NOR so it must have been 5H or 6H, and the centric translocation was 4HS.2BL. Following selfing, homozygotes of four translocations were selected. The experiment suggests that in vitro culture conditions are conducive for major genome rearrangements in wheat-barley hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Recombination of gliadin genes of chromosome 1D in the common wheat hybrid carrying the introgression from Aegilops cylindrica

Recombination of co‐gliadin components of the allele ‘_/’ at the Gli‐D1 locus and the allele at the respective locus from Aegilops cylindrica was revealed in the common wheat hybrid carrying the introgression from Ae. cylindrica. Gliadins of single seeds from F₂ plants of the cross between the winter common wheat variety ‘Albatros Odesskii’ and the common wheat line carrying the introgression from Ae. cylindrica were analyzed by acid polyacrylamide gel electrophor‐esis. The average frequency of recombination detected between the gliadin components was 0.35%.     

Somatic in vitro culture response of Lolium perenne L.: genetic effects and correlations with anther culture

Summary Genotypic effects on callus induction and plant regeneration in callus, suspension and protoplast culture, and their correlations with both phenotypic and GCA-values for anther culture response, were studied using 21 genotypes of perennial ryegrass. Differences between genotypes accounted for approximately 40% of the total variation for callus induction and initial callus growth, and 59 and 83% of the variation in callus culture for regeneration percentage and percentage of green plants. Effects of genotypes were less pronounced in suspension culture, where suspensions from the same genotype often behaved differently. Some suspension cultures retained their capacity for green plant regeneration for almost two years, repeatedly producing 80–100% green regenerants during this period. Genotypes with high regeneration percentage and a large proportion of green plants from callus culture were also superior in suspension culture for both regeneration performance and longevity. Regeneration percentage and percentage of green plants were uncorrelated, and probably under different genetic control. While capacity for green plant formation from the different genotypes showed no correlation between anther culture and somatic in vitro culture, a positive correlation was observed between the regeneration percentages in somatic in vitro culture and anther culture (r=0.44^*–0.85^***), suggesting some common genetic control of the two systems.     

In vitro techniques for selecting wheat (Triticum aestivum L.) for Fusarium-resistance. II. Culture filtrate technique and inheritance of Fusarium-resistance in the somaclones

Summary Calli of resistant, intermediary and susceptible wheat (Triticum aestivum L.) varieties were selected using culture filtrates of Fusarium graminearum and F. culmorum and the regenerants were evaluated for resistance up to R₃. Czapek-Dox broth medium was inoculated with mycelia of Fusarium isolates and incubated for 2–6 weeks. Filtrates were added to MS callus growing medium, then 5 weeks-old calli were transferred onto this medium (MST) for 4–5 weeks. MST containing 30% filtrate was found to be suitable for selection. Resistant calli were transferred again to fresh MST for further two selection cycles. The surviving calli produced less fertile regenerated lines (R₀) than the non-selected ones. Among 18 R₁ lines tested for Fusarium-resistance in the seedling stage by artificial inoculation in the greenhouse, two (11.1%) were significantly more resistant, one (5.6%) was more susceptible than the original cultivar and the rest (83.3%) behaved similarly to the donor plants. Among unselected R₃ lines of three varieties, practically the same number of resistant plants were found as among the related selected ones. When the R₃ selfed generations obtained through double-layer and culture filtrate selection techniques were tested for Fusarium-resistance, 35.7% of the lines were found to be more resistant than the original cultivars, none was more susceptible and 64.3% had a reaction similar to that of the source materials. Thus, inheritance of the disease reaction was not stable in all cases. Success of in vitro selection for Fusarium-resistance depended also on the genotype, and toxin analysis showed that although being effective, the selective media contained deoxynivalenol only exceptionally. In selecting wheat for Fusarium-resistance in vitro, the culture filtrate technique proved better than the double-layer procedure.     

Estimation of genetic parameters for in vitro traits in wheat immature embryo cultures involving high X low regeneration capacity genotypes

Summary Estimates of gene actions were obtained for five in vitro traits of immature wheat (Triticum aestivum L.) embryo cultures from a cross of two wheat cultivars and the resulting reciprocal, F₁, F₂ and backcross populations. The contribution of additive gene effects to in vitro traits was not as important as the dominance gene effects. Epistatic gene effects were relatively more important than either additive or dominance gene effects. Of the individual types of digenic epistatic effects, the dominance x dominance estimates were relatively larger in magnitude for all in vitro culture traits measured. The maternal effect played a minor role in the inheritance of the in vitro studied traits since the difference among the reciprocal values was not significant. It is shown from the generation mean method that epistasis played a major role in the inheritance of most of the traits under study. The negative values of additive and dominance genetic variance were estimates of zero. Heritability estimates, in broad sense, were relatively high for the in vitro studied traits. In some cases, heritability estimates in broad and narrow senses are almost equal since the estimation of dominance genetic variance led to negative values. According to the results of the gene effects, dominance and epistasis were important for the shoot formation trait. Selection would be effective among the isolated genotypes on individual basis.     

Gli-D t T1 and a novel γ-gliadin gene in Aegilops tauschii

Genetic studies of F₂ progeny from a cross between Aegilops tauschii accessions AUS18913 and CPI110856 revealed that Gli‐D^tT1 is tightly linked with a γ‐gliadin gene at the Gli‐D^t7 locus located on the short arm of chromosome 1D. Acid‐polyacrylamide gel electrophoresis (A‐PAGE) used to examine the gliadins of this population revealed two genes that encode a ω‐gliadin (T₁) protein and a novel γ‐gliadin protein. The ω‐gliadin (T₁) protein and the novel γ‐gliadin protein cosegregated, while having 0.69 cM recombination with the other locus (Gli‐D^t1). N‐terminal sequences were used to further classify the proteins studied.     

Location of genes for high grain protein percentage and other quantitative traits in wild wheat Triticum turgidum var. dicoccoides

Summary The genetic control of grain protein percentage (GPP) in the wild tetraploid wheat, Triticum turgidum var. dicoccoides, was determined by crossing four accessions of this taxonomic variety with durum cultivar Inbar, and analyzing the parents, F₁ and F₂ populations. Reciprocal crosses indicated no cytoplasmic effect on GPP. The F₂ variation was continuous in all crosses, showing no transgressive segregation. However, crosses between different accessions of var. dicoccoides showed transgressive segregation indicating the presence of different genes for high GPP in these accessions. Grain protein percentage was mostly codominant with high GPP, showing either no dominance, or a weak dominance. Heritability coefficients (broad sense) ranged from 0.30 to 0.53. Correlation coefficients between GPP and yield components were usually significantly negative, with the exception of the number of spikelets per spike, and in some crosses, grain weight.The number and chromosomal location of genes coding for high GPP were determined by the association between GPP and 27 markers (23 morphological and 4 biochemical markers). For this purpose, the genetic control of these markers, their linkage groups and chromosomal location were studied. At least four loci for high GPP that segregated in the F₂ populations are suggested: one on chromosome arm 1AS, marked by the black glume gene (Bg); one on 1BS, marked by the HMW gliadin locus Gli-B1; one on group 5, marked by the genes for beaked glume (Bkg) and toothed palea (Tp); and one on group 7, marked by the kinky neck gene (Kn). The relationship between GPP and several yield components was studied in a similar manner. In general, loci of markers that correlated positively with high GPP were not correlated with a decrease in yield components. Moreover, several loci of var. dicoccoides were associated with an increase in yield components.The utilization of markers for chromosomal location of genes coding for quantitative traits is compared to the technique of aneuploid analysis, commonly used in wheat. The significance of the above findings for breeding is discussed.