Ribosomal DNA spacer-length polymorphisms in three samples of wild and cultivated barleys and their relevance to the origin of cultivated barley

This study was conducted to provide additional data for evaluating two important issues surrounding the origin of cultivated barley: (i) the level of genetic diversity of the two‐rowed wild barley from Tibet, and (ii) the distribution of rDNA allele 104 in wild and cultivated barleys in the Occidental region. A total of 198 accessions consisting of three distinct samples were used: 82 entries of two‐rowed wild barley from Tibet, 57 accessions of two‐rowed wild barley from 8 countries with a broad range of representation of two‐rowed wild barley in the world, and 59 landrace accessions from four countries representing a part of the barley‐growing areas in the Middle East. These were assayed for rDNA spacer‐length variants (slvs). In all, 27 rDNA space length pheno types were detected, from which 10 slvs were identified as alleles at the two rDNA loci. The two‐rowed wild barley samples from Tibet had the lowest level of genetic variation as evaluated by rDNA polymorphism. Together with results of previous studies, the two wild forms (two‐rowed and six‐rowed) from Tibet could not account for the large genetic diversity observed in the cultivated barley of this region, suggesting that Tibet is unlikely a centre of origin for cultivated barley. In samples from the Occidental region, allele 104 of Rm2 was very rare in wild barley, but occurred at the highest frequency in cultivated barley, while the reverse is the case for allele 107, which is consistent with previous results. The implications of such a contrasting distribution of these rDNA alleles between wild and cultivated barleys in the origin and evolution of cultivated barley were discussed.     

Diversity and geographical distribution of seed lipoxygenase‐1 thermostability types in barley

To understand the diversity in the thermostability of the seed lipoxygenase‐1 (LOX‐1), 1040 cultivars of worldwide barley (Hordeum vulgare ssp. vulgare) genetic resources were investigated. The relative thermostability of LOX‐1 (LOX‐RTS) in these lines showed a bimodal frequency distribution and these lines were categorized into the high and low thermostability types (H‐type and L‐type, respectively). The H‐type lines predominated in the wild progenitor, ssp. spontaneum. The geographical distribution of these types in the cultivars was surveyed. The frequencies of the H‐ and L‐types were almost equal to one another in southwestern Asia. The occurrence of the H‐type predominated in eastern Asia and Africa, whereas in Europe and Turkey, the L‐type did. The predominance of the L‐type in Europe and Turkey can be understood through the hypothesis that the Fertile Crescent domestication contributed the majority of diversity in Europe. The uneven geographical distribution of the LOX‐1 thermostability types in the cultivars may reflect a polyphyletic origin of barley.     

Genetic characterization of wild barley populations (Hordeum vulgare ssp. spontaneum) from Kazakhstan based on genome wide SNP analysis

The wild ancestral form of barley, Hordeum vulgare ssp. spontaneum, is a valuable source for gene enrichment of cultivated barley. The purpose of this work was to study the area of distribution as well as the extent and structure of genetic variation of wild barley populations grown in Kazakhstan. It was found that distribution of wild barley populations in Kazakhstan is restricted to the most southern province. A genome wide single nucleotide polymorphism (SNP) analysis was performed in order to study the level of the genetic diversity in 96 accessions representing 14 wild barley populations from Kazakhstan and 25 accessions from the Middle East which is the center of diversity of this subspecies. The oligonucleotide pooled assay was used to genotype 384 SNPs distributed throughout the genome. In total 233 polymorphic SNPs were selected for further statistical analysis. The level of genetic diversity of wild barley populations from Kazakhstan was predictably narrower (He = 0.19 ± 0.01) in comparison with wild barley samples from the Middle East (He = 0.29 ± 0.01). The results suggested that H. vulgare ssp. spontaneum populations in Kazakhstan probably represent a recent spread of a limited number of plants from the primary distribution area and might be well adapted to winter low temperature.     

Random amplified polymorphic DNA (RAPD) variation in Fagopyrum tataricum Gaertn. accessions from China and the Himalayan region

The diversity among 52 landraces and cultivars of tartary buckwheat (Fagopyrum tataricum Gaertn.) and one accession of its wild ancestor, F. tataricum ssp. potanini Batalin, from diverse geographic origins was examined using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) markers. Eighteen primers produced a total of 240 fragments, of which 153 (63.75%) were monomorphic and 87 (36.25%) polymorphic bands. UPGMA-based pairwise Jaccard’s coefficient of similarity was used to deduce the relationships among 53 genetically diverse accessions. The similarity between cultivated tartary buckwheat accessions ranged from 0.61 to 1.00. Four distinct clusters were formed which corresponded well with the geographic distribution of the tartary buckwheat. Nepalese accessions showed maximum diversity followed by Chinese accessions. Tartary buckwheat accessions from the Himalayan region of northwestern India revealed a narrow gene pool. The wild buckwheat accession did not group with any of the three cultivated tartary buckwheat groups, and formed its own single-entry group. Genetic similarity (0.59) of Chinese buckwheat accessions with the wild ancestor reaffirmed that cultivated tartary buckwheat originated in the Yunnan province of northwestern China. Consistent with some earlier reports, our study demonstrated the usefulness of the RAPD technique for the characterization of plant genetic resources and assessment of diversity between species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Geographical differentiation and diallel analysis of seed dormancy in barley

Seed dormancy is one of the most important parameters affecting the malting process and pre-harvest sprouting in barley (Hordeum vulgare L.). Variation of seed dormancy in 4365 cultivated and 177 wild barley (ssp. spontaneum) accessions derived from different regions of the world was investigated in Okayama University, Kurashiki, Japan. Seed dormancy of each accession was estimated from their germination percentages at 0, 5, 10 and 15 weeks post-harvest after-ripening periods. All of the wild barley accessions showed less than 10% germination at 0 week after-ripening period. Level of seed dormancy in 4365 cultivated barley accessions showed a clear geographical differentiation. Seventy seven percent of Ethiopian accessions showed high germination percentages, while 86% of Japanese, Turkish and North African accessions showed low germination percentages at 0 week after-ripening period. A half diallel cross using eleven barley accessions with different level of dormancy revealed that seed dormancy was predominately controlled by additive gene effects. These results suggest that large genetic diversity for seed dormancy in barley is explained as different levels of additive accumulation of genetic factors. Barley varieties showing appropriate dormancy could be developed by crossing among barley germplasm accessions used in the present study.     

Variation in response to infection with Erysiphe graminis hordei and Puccinia hordei in some wild barley populations in a centre of diversity

Summary The Near East Fertile Crescent extending from Iran to Israel is the centre of origin of cultivated barley and a region of great genetic diversity in wild barley, Hordeum spontaneum C. Koch (syn. H. vulgare ssp. spontaneum (C. Koch Thell)). Wild barley accessions collected from different parts of this region were evaluated for their reactions to infection with three isolates of Erysiphe graminis hordei and two of Puccinia hordei. One culture of each pathogen was isolated in Israel and the others, either in Japan or the United States. Out of a total of 330 wild barley accessions collected from 14 sites in Iran, Turkey and Syria, only 18.8% were resistant to the Israeli culture, and 14.8% were resistant to a composite of the Japanese and American cultures of E. graminis hordei. Out of 105 accessions collected from six sites in Iran and Turkey, none was found to be resistant to the Israeli culture and 34.3% were resistant to the American culture of P. hordei. Considerable variation was observed both within and among sites for reactions to infection with different cultures of each of the two pathogens. The results of this study were compared with those of an earlier study involving wild barley accessions from Israel to illustrate the relative importance of different subregions in the Near East Fertile Crescent as sources of new genes for resistance to E. graminis hordei and P. hordei. Implications of these studies for in situ conservation of genetic diversity in wild barley are discussed.     

Mapping of Malt Endopeptidase, NADH Diaphorase and Esterase Loci on Barley Chromosome 3L

Isoelectric focusing (IEF) and immunoblotting were used to detect genetic variants of malt endopeptidase (MEP1), an enzyme related to malting quality in barley and coded by the CepB locus on barley chromosome 3 (= 3H). A variant was found in a Turkish accession of wild barley (Hordeum vulgare ssp. spontaneum). The self progeny of a hybrid between this accession and the barley cultivar ‘Clipper’ were analyzed for recombination between the CepB locus and other isozyme loci. The estimates of recombination linked the CepB locus to an NADH diaphorase locus (Ndh2), which in turn was linked to the seedling esterase complex (Est1, Est2, and Est4) situated near the distal end of the long arm of chromosome 3. The Ndh2 locus was independent of two other NADH dehydrogenase loci (Ndh3, Ndh5) which were mapped on barley chromosome 5 (= 1H) in relation to the three hordein loci (Hor1, Hor2, and Hor3).     

Analysis of the yield potential and yield components of F1 and F2 hybrids of crosses between wheat (Triticum aestivum L.) and spelt (Triticum spelta L.)

Summary Crosses of wild barley (Hordeum vulgare ssp. spontaneum and Hordeum vulgare ssp. agriocrithon) with Hordeum vulgare ssp. vulgare were used to select high yielding grain types under dryland Mediterranean conditions. No special difficulties were faced in making the crosses, in eliminating the brittle rachis genes from the grain types or in selecting 6-rowed types in crosses between 2-rowed wild barley and 6-rowed ssp. vulgare varieties. Brittle rachis genotypes, present in the segregating populations were used in developing self-reseeding permanent pastures for dry areas. The best selections were tested in seven trials during 1989–92 and some of them outyielded their parents and also the best improved check variety by 13–22%. Indications for transgressive segregation were obtained for grain yield, straw yield, total biological yield, harvest index and volume weight. The crude protein content of some of the selections was significantly higher than that of the checks. For breeding programs aiming at large seeds, special ssp. spontaneum lines should be used as parents. High grain yield was positively correlated with high straw yield, total biological yield, earliness in heading date, high harvest index and negatively with volume weight. It was concluded that unexploited useful genes, even when not directly observed in wild barley, could be transfered easily into high yielding genotypes by breeding.     

Utilization of exotic germplasm in Nordic barley breeding and its consequences for adaptation

Summary Utilization of exotic germplasm is one way to broaden genetic variation in breeding populations. This approach has recently been adopted in Sweden and Finland, where experimental barley populations has been established for research and pre-breeding purposes. The aim of the project is threefold: (1) to increase overall genetic diversity of Nordic barley breeding material; (2) to develop breeding material which possesses a high level of resistance for various barley diseases; and (3) to study effects of exotic germplasm on adaptation and agronomic performance. Both the Finnish and the Swedish barley populations include the same exotic material i.e. unadapted landraces from different parts of Asia and wild barley (Hordeum vulgare ssp. spontaneum) accessions. Locally adapted high-yielding barley lines were included in the populations. The establishment of these populations involved six crossing generations in order to promote recombination and enhance the break-up of linkage blocks. The paper discusses the third aim of the project. Studies on agronomic performance and adaptation showed that (1) agronomically valuable genotypes can be constructed through recombination using exotic germplasm for Nordic conditions, (2) that incorporation of exotic material is most successful when made in a local genetic base and (3) that exotic germplasm has an effect on adaptation.     

Molecular characterization and sequence polymorphism of the alcohol dehydrogenase 1 gene in Hordeum vulgare L.

Partial sequences (appr. 900bp) spanning from exon 2 to 4 of the gene encoding alcohol dehydrogenase 1 were obtained from four varieties of cultivated barley, Hordeum vulgare ssp. vulgare, and from 21 accessions of its wild progenitor, H. vulgare ssp. spontaneum. Sequence diversity was low and restricted to the non-coding intron sequences. In the 648–660bp long intron 3 both a microsatellite locus and a major duplication (153/156bp) were found. Sequence diversity in H. vulgare ssp. spontaneum was slightly higher than in cultivated barley. In general the Adh1 gene displays only limited polymorphism and is not appropriate for seeking correlations between molecular and eco-geographical data. This revised version was published online in July 2006 with corrections to the Cover Date.     

Variation in utilization efficiency and tolerance to reduced water and nitrogen supply among wild and cultivated barleys

Broad genotypic variation in the response to low soilmoisture and reduced nitrogen supply was found amongthe wild Hordeum spontaneum accessions and thelandraces and modern cultivars of H. vulgare ofdifferent geographic origin. Measurements at the endof vegetative growth in plants grown in soil culturesrevealed genotypically specific responses to the usedenvironmental factors. Cultivars and breeding linesfrom Syria and Ethiopian landraces combined bothdrought resistance and tolerance to low nitrogen. TheSyrian barleys were also distinguished by a highnitrogen utilization efficiency (NUE) under low Nnutrition. European cultivars indicated a pooradaptation to N shortage, but some of them wereresistant to soil drought. No stress resistant barleyswere found among the wild accessions and Sardinianlandraces. Genotypic differences in the relativevalues of NUE and water use efficiency were associatedwith low N-tolerance. Some Syrian selections,Ethiopian landraces and the modern German cv. Maresiwere found to be most drought resistant. Maintenanceof a relatively high photosynthetic activity of theuppermost leaves was associated with droughtresistance. As far as concerning with the vegetativegrowth phase, the modern Syrian germplasm andEthiopian landraces may be recommended as donors ofadaptative characters for local barley breeding.     

The transfer of a gene conferring resistance to scald (Rhynchosporium secalis) from Hordeum bulbosum into H. vulgare chromosome 4HS

Scald is a serious foliar disease that infects barley (Hordeum vulgare L.) causing reduced yields and adversely affecting quality. A means to combat the disease is to breed cultivars that possess genetic resistance. However, all known resistance alleles have so far originated from within the primary genepool of barley. This reliance on H. vulgare and H. vulgare ssp. spontaneum as resistance sources may encourage virulent forms of the pathogen to become established. To broaden the genetic base of cultivated barley and provide novel resistances to many diseases we have used a species from the secondary genepool of barley, H. bulbosum, in a resistance‐breeding programme. In this study we describe the development and trialling of a scald‐resistant recombinant line derived from a hybrid between H. vulgare and H. bulbosum. The scald resistance is simply inherited and located on the short arm of barley chromosome 4 (4HS).     

A novel barley scald resistance gene: genetic mapping of the Rrs15 scald resistance gene derived from wild barley, Hordeum vulgare ssp. spontaneum

Most genes for resistance to barley leaf scald map either to the Rrs1 locus on the long arm of chromosome 3H, or the Rrs2 locus on the short arm of chromosome 7H. Other loci containing scald resistance genes have previously been identified using lines derived from wild barley, Hordeum vulgare ssp. spontaneum. A single dominant gene conditioning resistance to scald was identified in a third backcross (BC3F3) line derived from an Israeli accession of wild barley. The resistance gene is linked to three microsatellite markers that map to the long arm of chromosome 7H; the closest of these loci, HVM49, maps 11.5 cM from the resistance gene. As no other scald resistance genes have been mapped to this chromosome arm, it is considered to be a novel scald resistance locus. As the Acp2 isozyme locus is linked to this scald resistance locus, at 17.7 cM, Acp2 is assigned to chromosome 7H. Molecular markers linked to the novel scald resistance gene, designated Rrs15, can be used in breeding for scald resistance.     

Comparison of European with West Asian and North African winter barleys in tolerance to boron toxicity

Three plastic-house experiments were conducted to compare the tolerance of European with West Asian and North African (WANA) winter barleys to boron (B) toxicity. Experiment I screened 24 winter barley entries with diverse origins. Experiment III tested 420 random accessions from seven European and seven WANA countries. Plants were screened in a soil mixed with boric acid (50 mg B/kg) and foliar B-toxicity symptom scores were recorded. Lower scores indicated higher B-toxicity tolerance. In Experiment II, five lines/varieties from each of the European and WANA groups were grown in pots with two soil B levels (0 and 25 mg B/kg). The West Asian landrace barleys had a lower mean B-toxicity symptom score than the European ones. The Syrian landrace variety normally grown in drier areas had a lower score than the Syrian landrace variety grown in wetter areas. Dry weights of the European and WANA groups were not different without adding B, but dry weight under 25 mg B/kg was lower for the European group than the WANA group. European accessions had a higher mean B-toxicity symptom score than the WANA accessions. Iranian and Afghan accessions had the lowest mean scores among countries. These results support the hypothesis that European winter barley varieties and accessions are less tolerant to B toxicity than those WANA accessions and varieties developed from local landraces. The lower B-toxicity tolerance could be a factor adversely affecting the performance of European winter barley varieties in the highlands of WANA. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genotypic variation in polyphenol content of barley grain

The polyphenol content in pearl barley, which is highly correlated to a browning reaction after heat treatment, was investigated using 1,347 cultivated barley varieties (H. vulgare) and two wild accessions (H. vulgare subsp. spontaneum) collected from different areas of the world. The polyphenol content in the cultivated barley shows a wide variation ranging from 0.19 to 0.75 mg/g with a nearly normal frequency distribution. The polyphenol content in the hulless varieties from Japan and Korea was low. On the other hand, the polyphenol content in wild barley was about two times higher than the average value recorded in cultivated barley. Based on HPLC analysis, five lowest-polyphenol content local varieties do not represent proanthocyanidin-free mutants. This revised version was published online in August 2006 with corrections to the Cover Date.     

Screening for resistance in the primary and secondary gene pool of barley against the root-lesion nematode Pratylenchus neglectus

Root-lesion nematodes of the genus Pratylenchus are significant pests in crop cultivation throughout many parts of the world. A study was initiated to determine the resistance of Hordeum vulgare and H. vulgare ssp. spontaneum (wild barley) against one major representative of the genus Pratylenchus , P. neglectus . A glasshouse test was first established. Barley seedlings were grown in 20 cm³ tubes filled with sand. Each plant was inoculated with 400 P. neglectus juveniles. After 12 weeks of cultivation nematodes were isolated from roots and sand using a misting chamber. The nematodes were counted under a microscope. A representative collection of 565 barley and wild barley accessions was tested in this way. The average number of nematodes per accession ranged from 350 to 12 000. In a verification experiment, 35 accessions with low and high infection rates were tested. This experiment identified a number of accessions with low infection rates. The perspectives for future breeding of barley cultivars resistant to root-lesion nematodes are discussed.     

Diversity in root architecture and response to P deficiency in seedlings of <Emphasis Type="Italic">Cucumis melo</Emphasis> L.

Breeding for more phosphorus (P)-efficient crops is one strategy to reduce the use of P fertilizers, thus mitigating the environmental and economic impacts of agriculture. Variation in root architecture and the response to P deficiency were studied in C. melo. Forty accessions representing genetic diversity within the species were screened for their root systems in normal and deficient P conditions at the seedling stage. Various parameters of P-uptake and P-use were analyzed in a subset of accessions at 40 days. Significant differences in root architecture were observed, with the taproot systems prevailing among the wild and exotic accessions, and more branched root systems in cultivated stocks. Moreover, differences in the plastic response of roots to P starvation were observed. Variation in different P-use and -uptake traits correlated with the root architecture. Within ssp. melo, the inodorus and flexuosus landraces had larger and more branched roots and more efficient P-uptake, thereby providing a close genepool for breeding. Within ssp. agrestis, conomon and momordica accessions can be sources of interest for the enhancement of variation in root architecture and P-use efficiency into cultivated melons. Therefore, the diversity observed within C. melo species could be useful in breeding P-efficient melon cultivars.     

Variation in developmental patterns of wild barley (Hordeum spontaneum L.) and cultivated barley (H. vulgare L.)

Summary Six wild barley (Hordeum spontaneum) accessions, from a diverse range of habitats, and two spring-cultivated barleys, were examined for variation in durations of development phases. The durations of the leaf initiation and spikelet initiation phases were longer and spikelet growth phases shorter, in wild than in cultivated barley. Across all wild and cultivated barleys the rate and duration of spikelet initiation were negatively correlated, but neither was related to the number of spikelets per spike. The number of spikelets was positively correlated with the number of leaves and the ratio of the number of spikelets to the number of leaves declined with increasing time to anthesis, indicating that each successive leaf was associated with a diminishing increase in the number of spikelets. The duration of culm elongation and final culm length were shorter in accessions of cultivated barley compared with wild barley. This paper also discusses the feasibility for increasing the number of spikelets per spike through breeding for genetic changes in lengths of pre-anthesis phases of development.Abbreviations ANOVA Analysis of variance - HV Hordeum vulgare - CE Culm elongation - DR Double ridge - HS Hordeum spontaneu - ° Cd Degree days     

Identification of Resistance Genes Against Powdery Mildew in Four Accessions of Hordeum Vulgare SSP. Spontaneum

Summary Four newly detected accessions of wild barley (Hordeum vulgare ssp. spontaneum) resistant to powdery mildew caused by Blumeria graminis f. sp. hordei were studied with the aim of finding the number of genes/loci conferring the resistance of individual accessions, the type of inheritance of the genes and their relationships to the Mla locus. F₂ populations after crosses between the winter variety ‘Tiffany’ and four wild barley accessions and use of microsatellite DNA markers were focused on the identification of individual resistance genes/loci by means of their chromosomal locations. In PI466495, one locus conferring powdery mildew resistance was identified in highly significant linkage with the marker Bmac0213. This location is consistent with the known locus Mla on chromosome 1HS. In the other three accessions the resistance was determined by two independent loci. In PI466197, PI466297 and PI466461, one locus was identified on chromosome 1HS and three new loci were revealed on chromosomes 2HS (highly significant linkage with Bmac0134), 7HS (highly significant linkage with Bmag0021) and 7HL (significant linkage with EBmac0755). Our prospective aim is identification of further linked DNA markers and the exact location of the resistance genes on the barley chromosomes.     

SSR-based genetic diversity assessment among Tunisian winter barley and relationship with morphological traits

For studying genetic diversity caused byselection for adaptation and end-use, 17microsatellites (SSR), representative ofthe barley genome, were used in 26 barley(Hordeum vulgare L.) accessions andcultivars in Tunisia. Theaccessions/cultivars originate fromdifferent geographic regions and are ofdifferent end-use. For the 15 polymorphicSSR, the mean number of alleles per locuswas 3.6 and the average polymorphisminformation content was 0.45. Clusteranalysis based on SSR data and onmorphological data clearly differentiatethe genotypes according to their type(local landraces vs. varieties), row-numberand end-use. The correlation between bothdiversity measures was highly significant(r = 0.25, p<10^-5) and thecorrespondence between the clustering basedon SSR and morphological data wasrelatively good. Our results show the largegenetic diversity of the Tunisian barleycultivars and the association of thisdiversity with adaptation traits.