Genetic diversity in European accessions of the Barley Core Collection as detected by isozyme electrophoresis

Genetic diversity in 79 European accessions of the Barley Core Collections was surveyed using isozyme electrophoresis. A total of 26 alleles were observed at the ten isozyme loci. All loci were polymorphic except Pgd-1 which was monomorphic. The comparison of the results with those of previous studies indicates that most of the alleles occurring in the European Barley are also observed in this set of the European Barley Core Collections. Only five alleles (Est-1 Al; Est-5 Ag, Te; Pgi-1 C and Ndh-2 B) were absent. Nine of 26 alleles were rare alleles, which were detected only in one or two accessions. Moreover, most of rare alleles were detected in 6-rowed winter barley. It is very important to include rare alleles for maximising the genetic variations in core collections. In the set of European Barley Core Collection, 6-rowed barley contained larger diversity than 2-rowed barley; winter type contained larger diversity than spring type. The cluster analysis separated 79 accessions into three major groups. Group I is more complex and comprised 2-rowed spring, 2-rowed winter and 6-rowed winter barley. In this group, 18 accessions in the cluster A and 14 accessions in the cluster B possessed identical genotypes as judged from the ten isozyme data. Principal coordinate analysis could not clearly separate the spring cultivars from the winter barley lines, as well as not separate 2-rowed from 6-rowed barley.     

Resistance to powdery mildew and barley yellow dwarf in perennial Triticeae species

Summary In a collaborative effort between the USDA-ARS and CAAS, 50 accessions each of perennial Triticeae held at the Forage and Range Research Laboratory, Logan, Utah, U.S.A. and the Institute of Crop Germplasm Resources, Beijing, People's Republic of China were exchanged. Eighty six and 85 accessions of these germplasms were screened for powdery mildew (caused by Erysiphe graminis DC) and barley yellow dwarf virus (BYDV) strain GPV, respectively. Fifty seven and 72 accessions had resistance to powdery mildew and BYDV, respectively. These materials are valuable genetic resources for breeding disease resistances in three of the major cereal crops — wheat, barley, and rye.Abbreviations BYDV barley yellow dwarf virus - ELISA enzyme-linked immunosorbent assay     

The Spanish barley core collection

Spanish barleys constitute a germplasm group of particular interest for breeding purposes, as Spain has been proposed as a possible centre of origin of the crop. The Spanish National Germplasm Bank (Banco Nacional de Germoplasma, BNG), holds a collection of about 2000 barley accessions, mostly landraces collected in Spain prior to extensive introduction of modern varieties. The objective of this work is to create a core collection of barleys representative of old barley genotypes grown in Spain. The core collection will be constituted by three groups of germplasm: successful old varieties (15); entries in common with previously existing barley core collections (15); and 2-row (8) and 6-row (122) entries from the BNG, for a total of 160 entries. Entries were allocated by stratified sampling in agro-ecological uniform zones of barley cultivation in Spain. Classification of agro-ecological regions for barley was based on historical yield records for Spanish provinces. The number of entries for each region was determined in proportion to the logarithm of historical barley acreage. Final choice of accessions within provinces tried to maximize the diversity and avoid duplications by looking at passport data, and to agronomic evaluation data available for a group of about 900 accessions.     

Genetic differentiation and geographical distribution of barley germplasm based on RAPD markers

Random amplified polymorphic DNA (RAPD) analysis was used to characterize barley germplasm genetic diversity. For the analysis 303 morphologically distinctive accessions were selected from the VIR germplasm collection, St. Petersburg, Russia and the MAFF Genebank, Tsukuba, Japan to represent the principal regions of barley cultivation. A total of 93 polymorphic bands scored from RAPD patterns were used to generate a genetic distance matrix, which was used in both cluster and principal coordinate analysis. Both analysis clearly separated barley cultivars and local populations into three distinctive groups, which evidently reflect different directions in evolution and geographical distribution of barley. The hierarchy of accessions clustering in the first group indicates the westward distribution of barley from West Asia to Europe and New World across Ethiopia and then Mediterranean region. The principal breeding trends based on spike morphology are also observed in this group. The second group is associated with eastward distribution of the crop and represents a unified genetic group, which consists of East Asian and Central Asian accessions. The third distinctive group identified is connected with the evolution and dissemination of hulless forms in Central Asia and the Caucasus region. The conformity of identified genetic groups and clusters with the global centers of crops diversity (gene centers) determined by Vavilov (1926) and modern ecogeographical classification of barley is discussed.     

Inferring geographic origin of barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L. subsp. <Emphasis Type="Italic">vulgare</Emphasis>) accessions using molecular markers

The USDA Agricultural Research Service (ARS) National Small Grains Collection (NSGC) has 207 landrace barleys obtained from a nursery grown in the Ukraine in 1930 by N.I. Vavilov, many of which have multiple resistance (MR) to disease similar to accessions from Ethiopia. The original collection locations of the accessions from the Vavilov nursery are unknown. The purpose of this study was to determine, using molecular markers, if the MR accessions of unknown origin from the Vavilov nursery are genetically related to MR accessions from Ethiopia. The genetic relatedness among susceptible accessions of unknown origin and among a selection of landrace accessions from the NSGC barley core subset was also assessed. Simple sequence repeat (SSR) and Diversity Array Technology PL (DArT) marker data were used to generate similarity matrices with Dice’s similarity coefficients. Cluster analysis from these results showed that the unknown-origin MR accessions grouped with accessions from Ethiopia. Susceptible accessions of unknown origin were genetically similar to accessions from western Asia, especially near the Caucasus, and from both southern and northern Europe. Based on similarity of marker profiles, some accessions from the core collection are likely duplicates. Future work will seek to identify the probable origin of the remainder of the unknown origin accessions and to more thoroughly characterize the genetic diversity within NSGC barley core subset.     

Identification of new sources for resistance to powdery mildew in H. spontaneum-derived winter barley lines

Twenty four barley lines derived from the F ₇generation of crosses between two winter barley cultivars and different accessions of Hordeum spontaneum Koch collected in Israel were tested against a set of ten European and five Israeli powdery mildew cultures, possessing virulence genes which completely match the spectrum of known mildew resistance genes. The comparison of reaction patterns justified the conclusion that new genes for resistance have been conferred from H. spontaneum which also differ from genes previously identified in other wild barley accessions from Israel. Participation in the expression of the resistance reaction of one of the two barley cultivars used in the development of the lines is well documented in some cases. The majority of the lines was found to be highly resistant against all or almost all European isolates, while various kinds of reactions were observed against the Israeli cultures. The effective transfer of novel mildew resistance from H. spontaneum into winter barley confirms similar results with spring barley, indicating natural wild barley populations in Israel as a significant gene pool for yet unexploited mildew resistance in barley.     

Phenotypic diversity in wild barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L. ssp. <Emphasis Type="Italic">spontaneum</Emphasis> (C. Koch) Thell.) accessions collected in Jordan

Wild barley, Hordeum vulgare L. ssp. spontaneum (C. Koch) Thell., is the progenitor of cultivated barley. Almost unanimously the center of diversity is considered to be in the Fertile Crescent of the Near East, where wild barley grows under a wide range of environmental and climatic conditions. Jordanian wild barley is expected to harbor genes useful for the improvement of cultivated barley, particularly those associated with tolerance to drought. This study evaluated 103 wild barley accessions collected from different areas of Jordan along with 29 cultivated barley genotypes for several morphological and agronomical traits. The Hordeum vulgare ssp. spontaneum C. Koch accessions were grouped into six populations according to the longitude, latitude, altitude, and rainfall zone of the collection site, and the cultivated barley in one population. The evaluation was conducted during the 2004–2005 growing season under field conditions in three locations in Jordan; namely, Khanasri, Ramtha, and Maru with 123.0, 222.9, and 429.2 mm annual rainfall, respectively. We used an unreplicated design with two systematic checks (the cultivars Rum and Mu’ta) each repeated 15 times. The results showed the existence of high variability among the Hordeum vulgare ssp. spontaneum C. Koch accessions for most of the traits, especially for plant height, tiller number, days to heading, days to anthesis, peduncle length, and peduncle extrusion. Plant height, earliness, peduncle length, and peduncle extrusion were found to be adaptive traits under drought conditions and several superior genotypes for each trait were identified. Genetic variation within population was much higher than between populations. Clustering of populations was according to their ecological geographical pattern.     

Genetic diversity of Acid-PAGE monomeric prolamins in cultivated hulless barley (Hordeum vulgare L.) from Qinghai–Tibet plateau in China

Naked barley, due to its favorable attributes such as high feed value, good human nutrition and easy processing, increasingly attracts people’s attention. Qinghai–Tibet Plateau is very rich in naked barley resources and has a long growing history. Genetic diversity of these cultivated naked barley is, however, poorly documented. This study analyzed the genetic diversity of monomeric prolamins (protein fraction corresponding to wheat gliadins) using the Acid -PAGE technique in eighty-six cultivated naked barley cultivars from Qinghai–Tibet Plateau. Extensive diversity was observed. A total of 43 different bands and 76 distinct patterns were found. Jaccard’s coefficient of similarity was calculated, and the accessions were divided into five main groups by cluster analysis using UPGMA. Differentiation among the populations from different collecting regions was investigated, based on the polymorphism of monomeric prolamins. The genetic diversity within the accessions from Tibet was slightly higher with an average diversity index of 0.29 than those from Sichuan. This study supports Tibetan Plateau is the diversity center of the cultivated naked barley and suggests that cultivated naked barley from Qinghai–Tibet Plateau is an important pool of variability that could be used in cereal breeding.     

Sources of powdery mildew resistance in a wild barley collection

A total of 1,383 accessions of wild barley (Hordeum vulgare ssp. spontaneum) held in the USDA-National Small Grains Collection, Aberdeen were screened for resistance to powdery mildew (Blumeria graminis f. sp. hordei) in the Czech Republic. They were subsequently tested at the seedling stage for reaction to selected powdery mildew pathotypes and simultaneously for their adult plant resistance in the field. One hundred and twenty-three accessions exhibited resistance to 22 pathotypes in the greenhouse tests (reaction type 2–3 or lower) as well as resistance to the natural population of this pathogen in the field in the Czech Republic during at least 2-year testing. These accessions represent large amount of promising sources in breeding barley for resistance to powdery mildew.     

Allelic diversity of a beer haze active protein gene in cultivated and Tibetan wild barley and development of allelic specific markers

The formation of haze is a serious quality problem in beer production. It has been shown that the use of silica elute (SE)-ve malt (absence of molecular weight (MW) ～14000 Da) for brewing can improve haze stability in the resultant beer, and the protein was identified as a barley trypsin inhibitor of the chloroform/methanol type (BTI-CMe). The objectives of this study were to determine (1) the allelic diversity of the gene controlling BTI-CMe in cultivated and Tibetan wild barley and (2) allele-specific (AS) markers for screening SE protein type. A survey of 172 Tibetan annual wild barley accessions and 71 cultivated barley genotypes was conducted, and 104 wild accessions and 35 cultivated genotypes were identified as SE+ve and 68 wild accessions and 36 cultivated genotypes as SE-ve. The allelic diversity of the gene controlling BTI-CMe was investigated by cloning, alignment, and association analysis. It was found that there were significant differences between the SE+ve and SE-ve types in single-nucleotide polymorphisms at 234 (SNP(234)), SNP(313), and SNP(385.) Furthermore, two sets of AS markers were developed to screen SE protein type based on SNP(313). AS-PCR had results very similar to those obtained by immunoblot method. Mapping analysis showed that the gene controlling the MW～14 kDa band was located on the short arm of chromosome 3H, at the position of marker BPB-0527 (33.302 cM) in the Franklin/Yerong DH population.     

Genetic diversity in Hordeum agriocrithon E. Åberg, six-rowed barley with brittle rachis, from Tibet

Twenty-two accessions of H. agriocrithon from Tibet were investigated for morphological and physiological characters, and genotypes combined alleles at the tightly linked Est1, Est2 and Est4 complex locus encoding esterase isozymes. H. agriocrithon was similar in plant type to Tibetan cultivated barley, but different from H. spontaneum. The accessions of H. agriocrithon were classified by their spike-awn types into vars. eu-agriocrithon, dawoense and paradoxon, with one exceptional accession of the awnless naked type similar to var. nuditonsum of cultivated barley. They were characterized by spring growth habit, susceptibility to Japanese race I of powdery mildew, and either of the same three esterase genotypes as those of cultivated barley from Tibet, Bhutan and Ladakh. The similarity suggested that the esterase complex locus on chromosome 3H is closely linked with some QTL for adaptation to Tibetan conditions. On the origin of H. agriocrithon, it might be concluded that natural hybridization between H. spontaneum and six-rowed barley occurred in northern parts of Afghanistan, Pakistan or India, and seeds of their segregants mixed into barley or wheat were brought up to Tibet, resulting six-rowed barley with brittle rachis.     

RAPD data do not support a second centre of barley domestication in Morocco

A RAPD analysis was conducted to clearify the mode of evolution of barley in the western Mediterranean basin. Twenty-nine barley accessions (Hordeum vulgare) and 13 populations of the wild progenitor (H. v. subsp. spontaneum) together with two outgroup species were analysed with seven RAPD primers, resulting in 101 polymorphic amplified fragments. Phenetic and cladistic analyses of the RAPD data resulted in trees indicating a monophyletic origin of cultivated barley, thus making a secondary domestication in Morocco unlikely. In Morocco spontaneous back mutation to wild type and crossing between wild barley (introduced from the E Mediterranean) and cultivated lines occurred, which gave rise to the local weedy forms of barley.     

Molecular analysis of the DNA polymorphism of wild barley (Hordeum spontaneum) germplasm using the polymerase chain reaction

Summary Hordeum spontaneum is the progenitor of cultivated barley (H. vulgare) and is an important source of genetic variation for barley breeding programs. The genetic diversity ofH. spontaneum in the Australian germplasm collection was investigated using the polymerase chain reaction with random and semi-random primers. This approach was found to be robust in respect of reaction conditions. Genetic dissimilarity values between genotypes were used to produce a phenogram of the relationships among the accessions using the unweighted pair group method with arithmetic mean. The largest divergence was observed among Israeli accessions, whereas the Turkish and Iranian samples clustered as distinct subsets, each apparently related to portion of the Israeli material. The results indicate that the genetic diversity of the wild barleys is broadly correlated with geographic distribution.     

Assessing Genetic Diversity of Chinese Cultivated Barley by STS Markers

To assess the genetic diversity among China’s cultivated barley, sequence tagged site (STS) marker analysis was carried out to characterize 109 morphologically distinctive accessions originating from five Chinese eco-geographical zones. Fourteen polymorphic STS markers representing at least one in each chromosome were chosen for the analysis. The 14 STS markers revealed a total of 47 alleles, with an average of 3.36 alleles per locus (range 2–8). The proportion of polymorphic loci per population averaged 0.84 (range 0.71–1.00); the mean gene diversity averaged 0.39 (range 0.28–0.49). The means of P and H_e were highest in the Yangtze reaches and Southern zone (P = 1.00; H_e = 0.46) and lowest (P = 0.71 He = 0.28) in the Yellow river reaches zone. The STS diversity in different zones is quite different from the morphology diversity. The STS variation was partitioned into 17% among the zone and 83% within the zone. Both cluster and principal coordinate analyses clearly separated the accessions into a dispersed group (mostly two-rowed barley with a lower mean GS value) and a concentrated group (mostly six-rowed barley with a higher mean GS value) according to the spike characteristic with only a few exceptions. The accessions from the Qinghai-Tibet plateau formed a distinctive subgroup and can be distinguished from the concentrated group. The role of Tibet in the origin and evolution of cultivated barley has been discussed.     

EST <Emphasis Type="Italic">versus</Emphasis> Genomic Derived Microsatellite Markers for Genotyping Wild and Cultivated Barley

Genetic variation present in wild and cultivated barley populations was investigated using two sources of microsatellite also known as simple sequence repeat (SSR) markers. EST-SSRs are derived from expressed sequences and genomic SSRs are isolated from genomic DNA. Genomic SSR markers detected a higher level of polymorphism than those derived from ESTs. Polymorphism information content was higher in genomic SSRs than EST-derived SSRs. This study showed that the EST-SSR markers developed in cultivated barley are polymorphic in wild and cultivated varieties and produced high quality markers. Ten of these functional markers were polymorphic across the accessions studied. EST markers indicated clearer separation between wild and cultivated barley than genomic SSRs. The EST-SSRs are a valuable source of new polymorphic markers and should be highly applicable to barley genetic resources, providing a direct estimate of functional biodiversity.     

Quantitative Trait Loci for Seed Dormancy in Wild Barley (Hordeum spontaneum C. Koch)

A quantitative trait locus analysis was carried out to unravel the genetic basis of dormancy in wild barley (Hordeum spontaneum) from Israel. Two accessions, Ashkelon and Mehola, from divergent environments were crossed to produce a mapping population. A linkage map was produced from the F₂ population, and F₄ seeds were used for germination experiments. Five quantitative trait loci (QTL) were detected for dormancy across the different germination experiments. These QTL were found on chromosomes 1, 2, 5, 6 and 7. The variation explained by each QTL varied between 8 and 25%. Ashkelon alleles increased the germination except for the QTL on chromosome 5. Three out of these five QTL co-locate with QTL found earlier in cultivated barley crosses, although this does not necessarily imply that they would be the same loci. The level of dormancy is much higher in wild barley than in cultivated barley and wild barley may have alleles that have not yet been utilised in breeding for optimally dormant barley.     

Evaluation of cultivated and wild barley for resistance to pathotypes ofPuccinia hordei with wide virulence

An isolate ofPuccinia hordei (ND89-3) originally collected in Morocco is virulent on most barley genotypes reported to possess resistance, except cultivar Estate (CI 3410), which possesses theRph3 gene and exhibits a low to intermediate level of resistance (infection type 12). Isolate ND89-3 possesses one of the widest virulence spectrums reported forP. hordei. Accessions ofHordeum vulgare (1,997 in total) andH. spontaneum (885 in total), mostly originating from the Mediterranean region and parts of North Africa, were evaluated with isolate ND89-3 at the seedling stage to identify new sources of leaf rust resistance. Fifty-eight accessions ofH. vulgare, and 222 accessions ofH. spontaneum exhibited low infection types to this isolate. Further evaluations of these resistant accessions with isolates ofP. hordei virulent forRph3,Rph7, andRph12 suggested that most of the resistantH. vulgare accessions possess theRph3 gene. Data suggested additional sources of effective resistance inH. vulgare are rather limited. FiveH. vulgare accessions and 167H. spontaneum accessions were identified as possible sources of new genes for leaf rust resistance. These accessions likely possess resistance genes that are different fromRph1 toRph12, or gene combinations thereof based on their reaction to four leaf rust isolates. Utilization of these accessions in barley breeding will broaden the germplasm resources available for genetic control ofP. hordei. North Dakota Agricultural Experiment Station Journal Series No. 2123. The U.S. Government right to retain a non-exclusive, royalty free licence in and to any copyright is acknowledged.