Phenotype/genotype associations for yield and salt tolerance in a barley mapping population segregating for two dwarfing genes.

Barley traits related to salt tolerance are mapped in a population segregating for a dwarfing gene associated with salt tolerance. Twelve quantitative trait loci (QTLs) were detected for seven seedling traits in doubled haploids from the spring barley cross Derkado x B83-12/21/5 when given saline treatment in hydroponics. The location of QTLs for seedling growth stage (leaf appearance rate), stem weight prior to elongation, and tiller number are reported for the first time. In addition, four QTLs were found for the mature plant traits grain nitrogen and plot yield. In total, seven QTLs are co-located with the dwarfing genes sdw1, on chromosome 3H, and ari-e.GP, on chromosome 5H, including seedling leaf response (SGa) to gibberellic acid (GA(3)). QTLs controlling the growth of leaves (GS2) on chromosomes 2H and 3H and emergence of tillers (TN2) and grain yield were independent of the dwarfing genes. Field trials were grown in eastern Scotland and England to estimate yield and grain composition. A genetic map was used to compare the positions of QTLs for seedling traits with the location of QTLs for the mature plant traits. The results are discussed in relation to the study of barley physiology and the location of genes for dwarf habit and responses to GA.

Wild barley: a source of genes for crop improvement in the 21st century?

The development of new barleys tolerant of abiotic and biotic stresses is an essential part of the continued improvement of the crop. The domestication of barley, as in many crops, resulted in a marked truncation of the genetical variation present in wild populations. This process is significant to agronomists and scientists because a lack of allelic variation will prevent the development of adapted cultivars and hinder the investigation of the genetic mechanisms underlying performance. Wild barley would be a useful source of new genetic variation for abiotic stress tolerance if surveys identify appropriate genetic variation and the development of marker-assisted selection allows efficient manipulation in cultivar development. There are many wild barley collections from all areas of its natural distribution, but the largest are derived from the Mediterranean region. The results of a range of assays designed to explore abiotic stress tolerance in barley are reported in this paper. The assays included; sodium chloride uptake in wild barley and a mapping population, effects for delta 13C and plant dry weight in wheat aneuploids, effects of photoperiod and vernalization in wild barley, and measurements of root length in wild barley given drought and nitrogen starvation treatments in hydroponic culture. There are examples of the use of wild barley in breeding programmes, for example, as a source of new disease resistance genes, but the further exploration of the differences between wild barley and cultivars is hampered by the lack of good genetic maps. In parallel to the need for genetic studies there is also a need for the development of good physiological models of crop responses to the environment. Given these tools, wild barley offers the prospect of a 'goldmine' of untapped genetic reserves.

The development and application of molecular markers for abiotic stress tolerance in barley.

This article represents some current thinking and objectives in the use of molecular markers to abiotic stress tolerance. Barley has been chosen for study as it is an important crop species, as well as a model for genetic and physiological studies. It is an important crop and, because of its well-studied genetics and physiology, is an excellent candidate in which to devise more efficient breeding methods. Abiotic stress work on cultivated gene pools of small grain cereals frequently shows that adaptive and developmental genes are strongly associated with responses. Developmental genes have strong pleiotropic effects on a number of performance traits, not just abiotic stresses. One concern is that much of the genetic variation for improving abiotic stress tolerance has been lost during domestication, selection and modern breeding, leaving pleiotropic effects of the selected genes for development and adaptation. Such genes are critical in matching cultivars to their target agronomic environment, and since there is little leverage in changing these, other sources of variation may be required. In barley, and many other crops, greater variation to abiotic stresses exists in primitive landraces and related wild species gene pools. Wild barley, Hordeum spontaneum C. Koch is the progenitor of cultivated barley, Hordeum vulgare L. and is easily hybridized to H. vulgare. Genetic fingerprinting of H. spontaneum has revealed genetic marker associations with site-of-origin ecogeographic factors and also experimentally imposed stresses. Genotypes and collection sites have been identified which show the desired variation for particular stresses. Doubled haploid and other segregating populations, including landrace derivatives have been used to map genetically the loci involved. These data can be used in molecular breeding approaches to improve the drought tolerance of barley. One strategy involves screening for genetic markers and physiological traits for drought tolerance, and the associated problem of drought relief-induced mildew susceptibility in naturally droughted fields of North Africa.

Using stable isotope natural abundances (delta 15N and delta 13C) to integrate the stress responses of wild barley (Hordeum spontaneum C. Koch.) genotypes.

To integrate the complex physiological responses of plants to stress, natural abundances (delta) of the stable isotope pairs 15N/14N and 13C/12C were measured in 30 genotypes of wild barley (Hordeum spontaneum C. Koch.). These accessions, originating from ecologically diverse sites, were grown in a controlled environment and subjected to mild, short-term drought or N-starvation. Increases in total dry weight were paralleled by less negative delta 13C in shoots and, in unstressed and droughted plants, by less negative whole-plant delta 13C. Root delta 15N was correlated negatively with total dry weight, whereas shoot and whole-plant delta 15N were not correlated with dry weight. The difference in delta 15N between shoot and root varied with stress in all genotypes. Shoot-root delta 15N may be a more sensitive indicator of stress response than shoot, root or whole-plant delta 15N alone. Among the potentially most productive genotypes, the most stress-tolerant had the most negative whole-plant delta 15N, whether the stress was drought or N-starvation. In common, controlled experiments, genotypic differences in whole-plant delta 15N may reflect the extent to which N can be retained within plants when stressed.

Phenotypic responses of wild barley to experimentally imposed water stress.

Responses to water stress within a population of wild barley from Tabigha, Israel, were examined. The population's distribution spans two soil types: Terra Rossa (TR) and Basalt (B). Seeds were collected from plants along a 100 m transect; 24 genotypes were sampled from TR and 28 from B. Due to different soil water-holding capacities, plants growing on TR naturally experience more intense drought than plants growing on B. In a glasshouse experiment, water was withheld from plants for two periods (10 d and 14 d) after flag leaf emergence. A total of 15 agronomic, morphological, developmental, and fertility related traits were examined by analysis of variance (ANOVA). Ten of these traits were significantly affected by the treatment. A high degree of phenotypic variation was found in the population with significant genotypextreatment and soil typextreatment interactions. Principal component analysis (PCA) was performed using combined control and stress treatment data sets. The first three principal components (pc) explained 88.8% of the variation existing in the population with pc1 (47.9%) comprising yield-related and morphological traits, pc2 (22.9%) developmental characteristics and pc3 (18.0%) fertility-related traits. The relative performance of individual genotypes was determined and water stress tolerant genotypes identified. TR genotypes were significantly less affected by the imposed water stress than B genotypes. Moreover, TR genotypes showed accelerated development under water deficit conditions. Data indicate that specific genotypes demonstrating differential responses may be useful for comparative physiological studies, and that TR genotypes exhibiting yield stability may have value for breeding barley better adapted to drought.

AFLP variation in wild barley (Hordeum spontaneum C. Koch) with reference to salt tolerance and associated ecogeography.

Thirty-nine genotypes of Hordeum spontaneum were selected from three geographically separated areas (southwestern, northern, and southeastern) of the Fertile Crescent. The lines were subject to AFLP (amplified fragment length polymorphism) analysis from which a similarity matrix was produced. A dendrogram of the data showed, with two exceptions, that the genotypes grouped together according to area of origin. This was confirmed by principal coordinate analysis in which the first principal coordinate separated the genotypes of the southwestern area from the other two areas, which in turn could be separated by the second principal coordinate. While genotypes from the same site of origin can exhibit very similar AFLP profiles, sharp genetic differences were detected between genotypes separated by relatively short distances. Thirty of the 39 genotypes were subjected to hydroculture salt tolerance tests. These were analysed for shoot Na+ content and carbon isotope composition (δ13C) after 4 weeks of treatment (100 mol∙m−3 NaCl). Shoot Na+ content and δ13C were highly correlated. Twelve AFLP markers were found to be associated with both shoot Na+ content and shoot δ13C and were also associated with site of origin ecogeographic data, particularly longitude. The most salt tolerant genotype came from Ilam in the southeastern area and the most salt sensitive genotype originated in the southwestern area. The 12 markers were partitioned into groups that showed significant associations within groups but no significant association between groups. In a multiple regression analysis, three AFLP markers, from separate groups, accounted for more than 60% of the variation for shoot Na+ content and δ13C. The results demonstrate the effectiveness of AFLP fingerprinting in genetic studies of complex traits at the wild species and (or) population level.

Detection of quantitative trait loci for agronomic, yield, grain and disease characters in spring barley (Hordeum vulgare L.).

Quantitative trait loci (QTLs) have been revealed for characters in a segregating population from a spring barley cross between genotypes adapted to North-West Europe. Transgressive segregation was found for all the characters, which was confirmed by the regular detection of positive and negative QTLs from both parents. A QTL for all the agronomic, yield and grain characters measured except thousand grain weight was found in the region of the denso dwarfing gene locus. There were considerable differences between the location of QTLs found in the present study and those found in previous studies of North American germ plasm, revealing the diversity between the two gene pools. Thirty-one QTLs were detected in more than one environment for the 13 characters studied, although many more were detected in just one environment. Whilst biometrical analyses suggested the presence of epistasis in the genetic control of some characters, there was little evidence of interactions between the QTLs apart from those associated with yield. QTLs of large effect sometimes masked the presence of QTLs of smaller effect.

Seed storage protein variation in Arachis species.

Seventy-two accessions, representing 22 species from sections Arachis, Erectoides, Extranervosae, and Triseminalae of the genus Arachis, were screened for seed storage protein polymorphism. Variation was detected between sections, between genome types, between species, and in some cases between different accessions of the same species or different seeds of the same accession. Arachis duranensis and one accession of A. cardenasii were found to have identical protein patterns. The greatest dissimilarity was found between species of the section Extranervosae and species of the section Triseminalae. Those of section Erectoides showed much similarity with some species of section Arachis. Protein polymorphism was shown to distinguish the two subspecies of A. hypogaea (fastigiata and hypogaea) in 27 of 28 cases. The seed protein profile of A. monticola was a combination of seed protein profiles from the two A. hypogaea subspecies. The relatedness between the various species was calculated and those that had the greatest similarity with A. hypogaea were A. spegazzinii and A. batizocoi.

Extraction and genetic control of two new water-soluble proteins of mature barley seed.

This paper describes the genetic control of two new water-soluble proteins in barley. Water-soluble proteins (WSPs) of mature barley seed form part of the albumin/globulin class of seed proteins. They can be extracted from hand-milled grain with water, though some WSPs are more efficiently extracted with a solution of 10 mM dithiothreitol. Polymorphisms for WSPs were detected in isoelectric focusing gels incorporating various ampholine combinations. Two new controlling genes (Wsp4 and Wsp5) have been identified and located using wheat/barley chromosome addition lines and barley doubled haploids. Wsp4 is located on chromosome 2 (2H), and Wsp5 was found to be tightly linked to Wsp2 on the long arm of chromosome 7 (5HL). Segregation of a sixth gene (Wsp6) is also described, but this has not been mapped. The results are discussed with respect to other previously mapped Wsp loci.

Molecular mapping of genes determining height, time to heading, and growth habit in barley (Hordeum vulgare).

A combination of random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) markers has been used to locate genes controlling important developmental characters in barley. The denso dwarfing gene has been mapped to the long arm of chromosome 3H. Stepwise multiple regression was also used to identify another region of the barley genome (on chromosome 7H), which contributed to variation in height. The denso locus was shown to be associated with delaying time to heading. A protein (WSP2) and an RAPD marker on barley chromosomes 5H and 6H, respectively, were also associated with time to heading. These results are discussed in relation to the genetic analysis of developmentally important traits and the development of dwarfing genes in barley breeding programs.

Genetic variation for waterlogging tolerance in the Triticeae and the chromosomal location of genes conferring waterlogging tolerance in Thinopyrum elongatum.

A number of Triticeae species were tested for tiller production, shoot dry matter production, and root penetration in waterlogged soil, and Thinopyrum elongatum and Elytrigia repens were shown to have better tolerance than wheat using these criteria. Tests of a number of wheat-alien amphiploids showed that there was at least partial expression of this exotic genetic variation in a wheat genetic background. The presence of chromosomes 2E and 4E of Th. elongatum was associated with a positive effect on root growth in waterlogged conditions. The positive effect of the 4E chromosome addition was mimicked by tetrasomic lines carrying extra doses of wheat homoeologues 4B and 4D, and it was concluded that the beneficial effect contributed by the presence of 4E was probably due to an increased dosage of group 4 chromosomes. However, the positive effect of adding chromosome 2E to wheat could not be reproduced by added doses of chromosomes 2A, 2B, or 2D, suggesting that this alien chromosome carries gene(s) for tolerance not present on its wheat homoeologues. This gene(s) was further located to the long arm of chromosome 2E by testing ditelosomic addition lines.

Statistical analysis of a linkage experiment in barley involving quantitative trait loci for height and ear-emergence time and two genetic markers on chromosome 4.

The quantitative traits height and ear-emergence date were analyzed in the F2 progeny of a cross between a tall winter barley cultivar (Gerbel) and a short spring barley cultivar (Heriot). The trait distributions were found to be related to the genotypes at two biochemical loci, ß-amylase (Bmy1) and water-soluble protein (Wsp3), which are known to lie on the long arm of chromosome 4. Linkages between each trait and the markers were investigated using normal mixture models. The two parental phenotypes and the heterozygote phenotype of Bmy1 were distinguishable so the model could be used directly to estimate linkage between Bmy1 and a quantitative trait locus (QTL) for height (Height). The Gerbel homozygote and heterozygote phenotype of Wsp3 could not be distinguished and the model was adapted accordingly. The proportion of plants requiring vernalization was consistent with control by two independent genes acting epistatically, and a normal mixture model based on a two-gene hypothesis was fitted to the distribution of ear-emergence date to estimate linkage between the marker loci and a QTL for ear-emergence date (Vrn1). The parameters of each model were the recombination fraction between the marker locus and the QTL and the means and standard deviations associated with each QTL genotype; these were estimated by maximum likelihood. The fitted distributions correspond well to those observed and the order of the loci along the chromosome is inferred to be Height - Vrn1 - Bmy1 - Wsp3, with Wsp3 being the most distal.

Characterization of rust-resistant wheat-Agropyron intermedium derivatives by C-banding, in situ hybridization and isozyme analysis.

Chromosome constitutions of three wheat-Agropyron intermedium derivatives were identified by C-banding analysis, in situ hybridization using biotin-labeled genomic Ag. intermedium DNA as a probe and isozyme analysis. Lines W44 and W52 were identified as 7Ai-2(7D) and 7Ai-2(7A) chromosome substitution lines carrying the same chromosome pair of Ag. intermedium. The alien chromosome was found to be homoeologous to group 7 based on C-banding, meiotic pairing and isozyme analyses. Line W49 was identified as a wheat Ag. intermedium chromosome translocation line. The breakpoint of the T2AS · 2AL-7Ai-2L translocation is located in the long arm at a fraction length of 0.62, and the transferred Ag. intermedium segment has a size of about 2.4 µm. Lines W44 and W52 expressed Ag. intermedium genes for resistance to leaf rust, stripe rust and stem rust, but only leaf rust resistance was expressed in W49. The results show that the leaf rust resistance gene(s), designated Lr38, is located in the distal half of the long arm of chromosome 7Ai-2, whereas the genes for resistance to stem rust and stripe rust are located either in the short arm or in the proximal region of the long arm of this chromosome.

Grain isozyme and ribosomal DNA variability in Hordeum spontaneum populations from Israel.

Grain isozyme and ribosomal DNA (rDNA) variability was examined in Hordeum spontaneum populations sampled from 27 geographical sites in Israel. Considerable phenotypic variability was observed with variants of ADH1, EST3, EST10, BMY1 and WSP detected, which are not available in the H. vulgare gene pool. Seven new rDNA phenotypes were detected in the H. spontaneum populations. Shannon's index of diversity was used to partition the total phenotypic variation into between and within population components. Most of the variation occurred between H. spontaneum populations. The distribution of both grain isozyme and rDNA phenotypes was non-random and correlated with a range of ecogeographical factors. In particular, the G phenotype of BMY1 was restricted to the Negev Desert and Dead Sea regions of Israel. Over 78% of the variation in the frequency of this particular phenotype could be explained by the number of rainy days per year and mean temperature in January. This suggests that variation at this locus or at loci linked to it may be of adaptive significance and of value in the introgression of genes controlling abiotic stress tolerance from H. spontaneum into the H. vulgare gene pool.

The inheritance of genetic markers in microspore-derived plants of barley Hordeum vulgare L.

Biochemical, molecular and morphological markers have been used to monitor the segregation of alleles at major gene loci in microspore-derived lines of four spring barley crosses and their parents. Significant deviations from the expected Mendelian ratios were observed for four of the ten markers studied in the cross. Distorted ratios were associated with loci located on chromosomes 4H and 6H. The differential transmission of alleles was in favour of the responsive parent (Blenheim) used in the anther culture studies. For the α-Amy-1 locus on chromosome 6H, the preferential transmission of Blenheim alleles was most pronounced in the haploid regenerants that were colchicine treated. These results are discussed in relation to the genetic control of androgenetic response in barley and with respect to the exploitation of another culture in barley improvement.

Water-soluble proteins of mature barley endosperm: genetic control, polymorphism, and linkage with ß-amylase and spring/winter habit.

Water-soluble proteins (WSP-2 and WSP-3) and ß-amylase (ß-AMY-1) were extracted from mature endosperms of 44 spring and 39 winter barley genotypes. The protein and enzyme isoforms were separated in isoelectric focusing gels with a pH gradient of 4-6.5. The Wsp-3 and ß-Amy-1 loci were located to chromosomes 4H using the wheat/barley chromosome addition lines. Segregation analysis of F2 and doubled haploid populations showed Wsp-2 and ß-Amy-1 to be tightly linked, with a map distance of 11 cMorgans. Isoforms of WSP-2 possessed similar pIs to that of WSP-3 and overlapping bands were observed in the gels. These bands segregated independently in F2 and doubled haploid populations, implying two unlinked genes. All three loci were found to be polymorphic: two alleles were detected at the Wsp-2 locus, three at Wsp-3 and two at ß-Amy-1. The frequency of alleles at all three loci was found to be different in winter and spring genotypes. Spring genotypes possessed a wider range of phenotypes than winter genotypes. Spring and winter genotypes could be distinguished on the basis of WSP-3 and ß- AMY-1 phenotypes. The linkage between Wsp-3 and ß-Amy-1 loci and genes controlling spring/winter habit on chromosome 4H is discussed. It is concluded that Wsp-3 and ß-Amy-1 can be used as genetic markers for spring/winter habit in barley genetic research and breeding.

Milling energy requirement of the aneuploid stocks of common wheat, including alien addition lines.

Aneuploid stocks, which included Triticum aestivum/alien, disomic, chromosome addition lines, wheat/alien, ditelosomic, chromosome addition lines, and the available aneuploids of "Chinese Spring" wheat, were used to locate genes that influence milling energy requirement (ME). Genes that affected ME were found on all seven homoeologous chromosome groups. The addition of complete wheat chromosomes 1B, 1D, 2A, 2D, 5B, 6B, 7B and 7D increased ME. Positive effects were also found in specific chromosome arms: 1BS, 2DS, 5AS, 5BS and 6BL. Wheat chromosome 3B conditioned low ME and the gene(s) responsible was located on the short arm. Other negative effects were attributed to wheat chromosome arms 4BL, 4DL, 5DS and 6DS. Alien chromosome additions that conferred high ME included 2H, 5H, 6H and 7H of barley, Hordeum vulgare and 2R, 2Rα, 4R, 4RL, 6R, 6RL and 7RL of rye, Secale cereale. Those that conferred a low ME included 1H (ch) of H. chilense, and 6u and 7u of Aegilops umbellulata, 5R and 5RS of S. cereale and 5R (m) and 5R (m)S of S. montanum. Although the control of ME is polygenic, there is a major effect of genes located on the short arms of homoeologous group 5 chromosomes.