Construction of immune lines with complex resistance to leaf rust and powdery mildew in common spring wheat cultivar Saratovskaya 29

Immune lines resistant both to leaf rust and to powdery mildew were constructed on the basis of common wheat cultivar Saratovskaya 29. Synthetic wheat Triticum timopheevii/Aegilops squarrosa (AAGGDD, 2n = 42) of Savov (Bulgaria) was used as a source of resistance genes. Using cytological analysis of BC2, we selected resistant plants (21") free from meiosis 1 (M1) defects. With these plants and continuous selection, BC8-BC9 immune lines were obtained. The lines were shown to carry new resistance genes differing from the known ones, and were proposed as donors of immunity to the diseases.     

A cytogenetic study of the blue-grain line of the common wheat cultivar Saratovskaya 29

The chromosome composition of the blue-grain line i:S29Ba of the cultivar Saratovskaya 29 was identified by cytological, GISH, and microsatellite analyses and C-banding. It was found that common wheat chromosome 4B of the cultivar Saratovskaya 29 was substituted with the Agropyron elongatum Host. chromosome carrying the gene for blue grain (s:S294Ag(4B)) during the construction of this nearly isogenic line. The blue-grain line was tested for productivity. The substitution of total chromosome 4B of the cultivar Saratovskaya 29 by Ag. elongatum chromosome 4 did not significantly affect the spike productivity parameters and grain quality with the exception of spike length (plus effect), spike density, and vitreousness (minus effects). The blue-grain line with s:S294Ag(4B) can be used in further studies associated with chromosome engineering in cereals and wheat breeding.     

Analysis of spike productivity traits in near-isogenic lines of the common wheat cultivar Saratovskaya 29 carrying alien marker genes

Six near-isogenic lines of the wheat cultivar Saratovskaya 29 carrying five marker genes from different species (Triticum compactum L., T. polonicum L., T. petropavlovskyi Udacz. et Migusch., Aegilops elongatum Host. and Secale cereale L.) were studied. It was shown that the introduced marker genes of taxonomic significance, C and P, have strong pleiotropic effects on quantitative traits of the spike productivity.     

Proteomic analysis of aneuploid lines in the homeologous group 1 of the hexaploid wheat cultivar Courtot

Three monosomic lines (MSLs) and three nullisomic lines (NSLs) of the homeologous group 1 and one euploid line of the bread wheat Triticum aestivum cultivar Courtot were used in a proteomic approach to investigate the effects of zero, one or two doses of chromosomes 1A, 1B and 1D on the amount of endosperm proteins. Polypeptides whose amounts changed significantly between each aneuploid line and the euploid line were identified using image analyses of two-dimensional gel electrophoresis patterns resulting from specific endosperm protein extractions. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry and electrospray ionization tandem mass spectrometry were also used for protein identification. Removing one chromosome or a chromosome pair allowed varying responses to be observed for the remaining endosperm protein genes. Compensation phenomena for the high molecular weight glutenin subunits (HMW-GS) were detected only in the MSLs. Subunits Bx7, By8 and Dy12 were the only HMW-GS overexpressed (from 152-737%) when chromosomes 1A or 1B or 1D were at hemizygous state. Thirteen new protein spots were detected only in the NSL1D, and seven were identified as HMW-GS analogs. These seven new spots may result from the expression of inactive genes. The HMW-GS were of significantly higher volume in MSLs, whereas the low molecular weight glutenin subunits and the gamma-gliadins were of lower volume in aneuploid lines. Most of the down-regulated proteins in the MSLs were storage proteins encoded at loci located on another chromosome pair. Complex regulations between chromosomes and loci of the homeologous groups 1 and 6 in bread wheat are discussed.     

The inheritance of endosperm storage proteins by the line of the Saratovskaya 29 cultivar of common wheat from its parental forms

We ran a comparative analysis of storage proteins (gliadins, high(HMW) and low-molecularweight (LMW) glutenins, puroindolines, and exogenous pest alpha-amylase inhibitors) in the Saratovskaya 29 cultivar line from the collection of a genetic engineering laboratory, its parental forms (Albidum 24 and Lutescens 55/11), and distant ancestors (Poltavka, Selivanovskiy Rusak, Sarroza, and tetraploid Beloturka). It was confirmed that the allelic states of storage proteins in the Gli-1, Gli-2 and Glu-1 loci originate from ancestral forms from the collection of the Vavilov Institute of Plant Industry. Moreover, new alleles were found in Lutescens 55/11 (Glu-A1a) and Selivanovskiy Rusak (Glu-B1b) cultivars from the collection of the Institute of Cytology and Genetics. A new allelic state, Ha, was observed in the loci of the Poltavka cultivar as a soft-grain cultivar, and the ha allele was found in the hard-grain Albidum 24 and Sarroza cultivars. It was found that the expression rate of exogenous pest alpha-amylase inhibitors in the Saratovskaya 29 cultivar line is lower than that of ancestral cultivars (Albidum 24, Sarroza, Poltavka, and Beloturka). Such inhibitors are absent in the paternal form Lutescens 55/11. A high expression rate of protein pest inhibitors for exogenous α-amylases and puroindolines was observed in the Poltavka cultivar. The allelic composition of Glu-1 loci was newly studied in the Sarroza cultivar, which has some promising features. The Saratovskaya 29 cultivar line, on the basis of which a wide range of diverse lines were created in the Institute of Cytology and Genetics, is isogenic for all of the studied traits.     

Characterization of puroindolines in the control of endosperm texture in common wheat lines with substitutions of homeologous group-5 chromosomes

The genetic control of grain hardness and its association with the specific friabilin content on starch granules of common wheat cultivars and lines with intervarietal substitutions of homeologous group-5 chromosomes were studied. A significant correlation was revealed between the technological parameters of grain hardness (mean size of flour particles) and the specific content of puroindolines on the starch surface estimated in terms of starch doses. The results obtained allowed the method of starch doses to be used to identify soft and hard wheat cultivars and lines based on an analysis of a single grain. The biochemical analysis confirmed the previously obtained estimates of flour-grinding properties of wheat cultivars and substitution lines and allowed specific genotypes to be characterized according to the composition of puroindolines. The influence of chromosomes 5D and 5A of donor wheat cultivars on the activity of the Ha loci of recipient cultivars was revealed and found to be associated with the composition of PIN products and with the expression of the Pina-D1 and Pinb-D1 genes.     

The effect of additions ofAegilops longissima chromosomes on grain protein in common wheat

Summary The effect of various chromosomes ofAegilops longissima when added to the common wheat cultivar Chinese Spring was evaluated at two levels of nitrogen fertilization for absolute and relative amount of protein in the grain. All the added chromosomes ofAe. longissima increased protein percentage: protein increase by chromosomes D, C and A averaged 3.8% while that by chromosomes F, E, G and B averaged 1.7%. Addition lines F, D and C had a significantly higher protein weight per grain. On the other hand, lines A, E and G had reduced grain protein weight per grain as compared with that of Chinese Spring. Line C carries the HMW glutenin and some of the gliadin subunits ofAe. longissima. The effect of this line, however, and obviously that of the other lines on protein content was through genes controlling the level of storage protein rather than through genes that code directly for these proteins. Nitrogen fertilization affected protein content and the relative amount of the various protein fractions in a similar manner in every addition line. When high levels of nitrogen fertilization were compared to low ones, the relative amount of the HMW glutenins remained constant while that of HMW gliadins increased and that of the LMW subunits decreased. In contrast to the nitrogen effect, increase in protein content by the addition oflongissima chromosomes did not change the relative amounts of the various protein fractions.The paper is based on a portion of a dissertation to be submitted by A.A.L. in partial fulfillment of the PhD requirements in the Feinberg Graduate School, The Weizmann Institute of Science, RehovotThe Marshall and Edith Korshak Professor of Plant Cytogenetics     

Transfer of grain softness from 5U-5A wheat-Aegilops triuncialis substitution line to bread wheat through induced homeologous pairing

Journal of Plant Biochemistry and Biotechnology - Bread wheat sustains genes for grain softness on “Ha” locus of short arm of 5D chromosome. Pina-D1 and Pinb-D1 alleles of...     

Features of crossability, haploidy and polyembryony in hybrid combinations between common barley Hordeum vulgare L. (2n = 14) and wheat-rye substitution lines Triticum aestivum L., cultivar Saratovskaya 29/Secale cereale L., cultivar Onokhoiskaya

The role of individual chromosomes of rye in the manifestation of crossability and seedling development in hybrid combinations between common barley Hordeum vulgare L., cultivar Nepolegayushchii (2n = 14) and five wheat-rye substitution lines Triticum aestivum L., cultivar Saratovskaya 29/Secale cereale L., cultivar Onokhoiskaya (2n = 40 wheat + 2 rye chromosomes). Crossability, which was measured by two parameters--frequency of set grains and frequency of grains with embryos--was shown to be significantly affected by each of the five rye chromosomes examined: 1R, 2R, 3R, 5R, and 6R; the development of barley haploids was affected by rye chromosomes 1 R, 3R, and 5R. We were the first to demonstrate that polyembryony could be induced by mutual effects of barley cytoplasm and rye chromosome 1R. Possible mechanisms controlling the development of haploids and twins in hybrid combinations H. vulgare x T. aestivum/S. cereale are discussed. The conclusion is drawn that hybrid combinations between common barley and wheat-rye substitution lines can serve as new models for studying incompatibility mechanisms in distant crosses and genetic control of parthenogenesis.     

Recent advances in the genetics underlying wheat grain protein content and grain protein deviation in hexaploid wheat

Wheat is one of the most important global crops and selection for better performance has been ongoing since ancient times. As a quantitative trait controlled by the interplay of several genomic loci and under the strong influence of the environment, grain protein content (GPC) is of major interest in breeding programs. Here, we review the most recent contributions to the genetics underlying wheat GPC and grain protein deviation (GPD, representing the relationship between grain protein content and yield), together with the performance of genomic prediction models characterizing these traits. A total of 364 significant loci related to GPC and GPD are positioned on the hexaploid wheat genome, highlighting genomic regions where significant independent QTL overlap, with special focus on two regions located on chromosomes 3A and 5A. Some of the corresponding homoeologous sequences co-locate with significant independent QTL reported on the B and D subgenomes. Overlapping independent QTL from different studies are indicative of genomic regions exhibiting stability across environments and genotypes, with promising candidates for improving grain quality.     

Genetic architecture of grain protein content in wheat

Studies on identification and localization of quantitative traits for grain protein content (QGpc-loci) on chromosomes in Triticum aestivum and Triticum durum are reviewed. Association of QGpc with various traits of morphology, physiology, adaptation and tolerance to abiotic and biotic stress is shown. Genetic and environmental QGpc contexts that should be taken into account when using molecular markers in breeding for the grain protein content are considered.     

Genetic architecture of grain protein content in wheat

Studies on identification and localization of quantitative traits for grain protein content (QGpc-loci) on chromosomes in Triticum aestivum and Triticum durum are reviewed. Association of QGpc with various traits of morphology, physiology, adaptation and tolerance to abiotic and biotic stressors is shown. Genetic and environmental QGpc contexts that should be taken into account when using molecular markers in breeding for the grain protein content are considered.     

Molecular genetic analysis of grain protein content and flour whiteness degree using RILs in common wheat

Grain protein content (GPC) and flour whiteness degree (FWD) are important qualitative traits in common wheat. Quantitative trait locus (QTL) mapping for GPC and FWD was conducted using a set of 131 recombinant-inbred lines derived from the cross ‘Chuan 35050 × Shannong 483’ in six environmental conditions. A total of 22 putative QTLs (nine GPC and 13 FWD) were identified on 12 chromosomes with individual QTL explaining 4.5–34.0% phenotypic variation. Nine QTLs (40.9%) were detected in two or more environments. The colocated QTLs were on chromosomes 1B and 4B. Among the QTLs identified for GPC, QGpc.sdau-4A from the parent Shannong 483 represented some important favourable QTL alleles. QGpc.sdau-2A.1 and QFwd.sdau-2A.1 had a significant association with both GPC and FWD. The markers detected on top of QTL regions could be potential targets for marker-assisted selection.     

The effect of rye chromosomes on callus induction and regeneration in callus cultures of immature embryos of wheat-rye substitution lines, Triticum aestivum L. cultivar Saratovskaia 29/Secale cereale L. cultivar Onokhoiskaia

The effect of individual rye chromosomes on the induction of callus and the character of its regenerating capacity was studied with cultured immature embryos of wheat-rye (Triticum aestivum L. cv. Saratovskaya 29-Secale cereale L. cv. Onokhoiskaya) substitution lines. The genotypic diversity of the substitution lines proved to significantly affect variation of parameters characterizing the major types of callus cultures, that is, frequencies of embryogenic calli, which are capable of shoot regeneration, and of morphogenic calli, which produce root structures. Functioning in the genotypic background of common wheat cultivar Saratovskaya, chromosomes 2R and 3R of rye cultivar Onokhoiskaya stimulated significantly the induction of embryogenic callus highly capable of shoot regeneration. Rye chromosome 2R present in place of chromosome 2D in the common wheat genome suppressed the induction of callus producing root structures. Rye chromosomes 1R and 6R suppressed the induction of embryogenic callus capable of shoot regeneration.     

Mapping QTLs for grain hardness and puroindoline content in wheat ( Triticum aestivum L.)

Genes for puroindoline-a (Pin-a), puroindoline-b (Pin-b) and grain-softness proteins (GSP) have been shown to be linked to the dominant Ha locus responsible for the soft texture of the grain. Though linkage has been demonstrated of the puroindoline genes to the Ha locus, there is no clear evidence that puroindoline content is the product of the gene Ha. A segregating population of 115 recombinant inbred lines (RILs) originating from a cross between the hexaploid Synthetic wheat ( Triticum durum x Aegilops tauschii, W 7984) and the cultivar 'Opata' (M 85) was studied in two different experimental years to detect Quantitative Trait Loci (QTLs) for three traits: grain hardness (Hard), puroindoline-a (Pin-a) and puroindoline-b (Pin-b) contents. The detection of QTLs was performed using marker linear regression. Negative correlation coefficients (-0.86 and -0.80) were identified between grain hardness and puroindoline content (a and b, respectively) on data obtained in 1996. Results obtained in 1999 confirmed the negative correlation between Hard and Pin-a (-0.73); however a positive correlation coefficient was found with Pin-b content (0.41). Total phenotypic variation explained by each QTL was calculated (R2). For each of the Hard, Pin-a and Pin-b traits one major QTL was detected on the short arm of chromosome 5D, located close to the mta9 allele (puroindoline-a). For the first year (1996) the QTL in this region explained around 63% of the phenotypic variability in grain hardness, 77% in Pin-a and 45% in Pin-b contents. These values were confirmed in trials carried out in 1999 with a R2 value of 0.71, 0.72 and 0.25 for Hard, Pin-a and Pin-b, respectively. In 1996 and 1999 a second major QTL was detected for grain hardness on the long arm of the same chromosome. Present results indicate that it cannot be definitely concluded that puroindoline content represents a linear explanation for variations in grain hardness.     

Epistasis and genotype-by-environment interaction of grain protein content in durum wheat

Parental, F(1) , F (2) , BC (1) and BC (2) generations of four crosses involving four cultivars of durum wheat (Triticum durum Desf.) were evaluated at two sites in Tunisia. A three-parameter model was found inadequate for all cases except crosses Chili x Cocorit 71 at site Sidi Thabet and Inrat 69 x Karim at both sites. In most cases a digenic epistatic model was sufficient to explain variation in generation means. Dominance effects (h) and additive x additive epistasis (i) (when significant) were more important than additive (d) effects and other epistatic components. Considering the genotype-by-environment interaction, the non-interactive model (m, d, h, e) was found adequate. Additive variance was higher than environmental variance in three crosses at both sites. The estimated values of narrow-sense heritability were dependent upon the cross and the sites and were 0%-85%. The results indicate that appropriate choice of environment and selection in later generations would increase grain protein content in durum wheat.     

Detection of QTLs for grain protein content in durum wheat

Grain protein content (GPC) of durum wheat (Triticum turgidum L. var. durum) is an important trait for the nutritional value of grain and for influencing the technological property of flour. Protein content is a quantitative trait negatively correlated with grain yield, thus increase in protein quantity usually results in yield reduction. This study was initiated to introgress alleles for high GPC from var. dicoccoides into durum wheat germplasm by the backcross inbred line (BIL) method and to identify molecular markers linked to high GPC alleles not associated with depressing effects on yield. The backcross line 3BIL-85 with high GPC and similar grain yield to the recurrent parent was backcrossed to Latino, and the generations F₂, F₃ and F₄ were evaluated for GPC and yield per spike (GYS) in three field trials. Three QTLs with major effects on GPC were detected on chromosome arms 2AS, 6AS and 7BL, identified by the markers Xcfa2164, XP39M37 ₍₂₅₀₎ and Xgwm577 _, respectively. Multiple regression analysis indicated that the three QTLs explained all the genetic variances of the trait. The high GPC parental line 3BIL-85 was not significantly different from the recurrent parent Latino for GYS, but the phenotypic correlation coefficient between GPC and GYS had negative values (from −0.02 to −0.28) in each trial, although it was statistically significant only in the F3 progeny trial. No co-located QTL for GYS was detected, excluding the hypothesis that the putative QTLs for GPC were indirect QTLs for low grain yield. The negative protein-yield response could be due to: (a) co-location of grain yield per spike QTLs with reduced phenotypic effects not detectable by the experimental design or statistical procedures, or to (b) opposite pleiotropic gene effects due to the major bio-energetic requirements for synthesis of protein then carbohydrates. Mapping loci by BILs should enable the production of near-isogenic lines in which the individual effects of each QTL can be examined in detail without confounding variations due to other putative QTLs. An erratum to this article can be found at     

Genetics of gliadins coded by the group 1 chromosomes in the high-quality bread wheat cultivar Neepawa

Summary The inheritance and biochemical properties of gliadins controlled by the group 1 chromosomes of the high-quality bread wheat cultivar Neepawa were studied in the progeny of the cross Neepawa x Costantino by six different electrophoretic procedures. Chromosome 1B of Neepawa contains two gliadin loci, one (Gli-B1) coding for at least six - or -gliadins, the other (Gli-B3) controlling the synthesis of gliadin N6 only. The map distance between these loci was calculated as 22.1 cM. Amongst the chromosome 1A gliadins, three proteins are encoded at the Gli-A1 locus whereas polypeptides N14-N15-N16 are controlled by a remote locus which recombines with Gli-A1. Six other gliadins are controlled by a gene cluster at Gli-D1 on chromosome 1D. Canadian wheat cultivars sharing the Gli-B1 allele of Neepawa were found to differ in the presence or absence of gliadin N6. The electrophoretic mobilities of proteins N6 and N14-N15-N16 were unaffected by the addition of a reducing agent during two-dimensional sodium dodecyl sulphate polyacrylamid-gel electrophoresis, suggesting the absence of intra-chain disulphide bonds in their structure.Research supported by a grant from the Commission of the European Communities, ECLAIR programme, Contract AGRE 0052