The resistance to leaf rust and powdery mildew of recombinant lines of barley (Hordeum vulgare L.) derived from H. vulgare × H. bulbosum crosses

A set of 23 recombinant lines (RLs) of barley ( Hordeum vulgare L.) derived from H. vulgare  ×  H. bulbosum L. crosses was inoculated with barley leaf rust ( Puccinia hordei ) and powdery mildew ( Blumeria graminis f.sp. hordei ) at the seedling stage to identify their levels and mechanisms of resistance. Eight RLs were studied further in glasshouse and field tests. All three barley parents ('Emir', 'Golden Promise' and 'Vada') were highly susceptible to powdery mildew and leaf rust isolates. Several RLs showed partial resistance expressed as high relative latency periods and low relative infection frequencies against leaf rust. This high level of partial resistance was due to a very high level of early aborting colonies without host cell necrosis. Several RLs showed hypersensitive resistance to some or all isolates. For powdery mildew, one RL was completely resistant to the CC1 isolate and had a hypersensitive resistance to the CO-02 isolate. Three RLs derived from 'Emir' were completely resistant to both powdery mildew isolates, and three more RLs tested in the field had higher levels of partial resistance than their parents. The results indicate that H. bulbosum contains major and minor gene(s) for resistance to leaf rust and powdery mildew that can be transferred to cultivated barley.     

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.     

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.     

Postulation of leaf-rust resistance genes in Czech and Slovak barley cultivars and breeding lines

Leaf‐rust resistance (Rph) genes in 61 Czech and Slovak barley cultivars and 32 breeding lines from registration trials of the Czech Republic were postulated based on their reaction to 12 isolates of Puccinia hordei with different combinations of virulence genes. Five known Rph genes (Rph2, Rph3, Rph4, Rph7, and Rph12) and one unknown Rph gene were postulated to be present in this germplasm. To corroborate this result, the pedigree of the barley accessions was analysed. Gene Rph2, as well as Rph4, originated from old European cultivars. The donor of Rph3, which has been mainly used by Czech and Slovak breeders, is ‘Ribari’ (‘Baladi 16’). Rph12 originates from barley cultivars developed in the former East Germany. Rph7 in the registered cultivar ‘Heris’ originates from ‘Forrajera’. A combination of two genes was found in 10 cultivars. Nine heterogeneous cultivars were identified; they were composed of one component with an identified Rph gene and a second component without any resistance gene. No gene for leaf rust resistance was found in 17 of the accessions tested. This study demonstrates the utility of using selected pathotypes of P. hordei for postulating Rph genes in barley.     

Identification of barley mutants in the cultivar 'Lux' at the Dhn loci through TILLING

TILLING is a reverse genetic strategy that allows screening for mutations in genes with known sequences in a plant mutant population. A TILLING population has been developed for the Danish barley variety 'Lux' ( Hordeum vulgare L.), by using sodium azide to induce mutations. Scoring of four visible phenotypic characters of barley seedling in reference to the parental cultivar 'Lux' in the M₃ plants showed over 3.5% lethality. A series of pool ratios of mixed DNA from mutant lines were tested and 10-fold pools appeared to be the practical mixing ratio for the detection of fragments in the 500–700 bp range. Two of the 13 known dehydrin genes, Dhn12 and Dhn13 , respectively, were examined and five independent missense mutations were obtained from a population of 9575 barley mutant plants. This corresponds to a mutation density of approximately one mutation every two and half million base pairs for these two genes. The mutant population of approximately 10 000 lines was screened for mutations in two genes in a short time due to high pooling ratio.     

Powdery mildew resistance in barley landrace material. I. Screening for resistance

A total of 4,681 accessions of Hordeum vulgare landrace material from Ethiopia, East Mediterranean, Near East, Nepal and China were sown in the field and subjected to the natural powdery mildew epidemic in Denmark. Apparently resistant accessions were selected. Selfed progeny from them were retested and reselected in subsequent years at four locations in Denmark. Finally, 16 promising donors of resistance were retained. They were characterized in the field and tested in the seedling stage for reaction to up to 72 different isolates of the powdery mildew fungus. The absence of the corresponding virulences in the Danish airborne powdery mildew population was ascertained in five years. The resistances in the 16 donors are apparently mutually different and from known sources of powdery mildew resistance in barley. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic resources of salt tolerance in wild Hordeum species

Salt tolerance was evaluated in 340 accessions of Hordeum, consisting of 41 brittle-rachis forms of Hordeum vulgare L. subsp. vulgare (H. agriocrithon) accessions, 154 H. vulgare L. subsp. spontaneum (H. spontaneum) accessions, and 145 accessions of ten other species or subspecies of wild Hordeum. Germination was carried out at concentrations of 171, 257, and 342 mM NaCl. The levels of salt tolerance for seed germination in wild Hordeum species were generally lower than those found by Mano et al. (1996) in cultivated barley; the NaCl tolerance level of the different species were as follows: H. agriocrithon > H. spontaneum > other wild Hordeum species. In addition, leaf injury index was used to assess tolerance at the seedling stage after treatment with 500 mM NaCl solution for four weeks. The levels of salt tolerance at the seedling stage in wild Hordeum species were generally higher than those found by Mano & Takeda (1995) in cultivated barley. Most wild Hordeum species showed high NaCl tolerance at the seedling stage and are considered good sources of germplasm for salt tolerance breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

Search for partial resistance against Puccinia hordei in barley landraces from the Fertile Crescent

A collection of 111 barley landraces from the Fertile Crescent was screened for resistance to barley leaf rust in the field and under controlled conditions. Large variation was observed for disease severity under field conditions. Accessions with high resistance because of hypersensitivity were identified. Also segregation was observed in some accessions, with individual plants showing hypersensitive reactions (IT ≤ 6). Partial resistance due to a reduction of infection in spite of a compatible infection was commonly found (19%). Resistance of 12 accessions selected for their low disease severity and high IT, was shown to be due to a prolonged latency period and increased percentage of early aborted colonies not associated with host cell necrosis. A high correlation was observed between the microscopic and macroscopic components of partial resistance.     

Genetic diversity and population structure of wild and cultivated barley from West Asia and North Africa

Population structure and relationships within and among 185 accessions of wild (Hordeum vulgare ssp. spontaneum) and cultivated barley (H. v. ssp. vulgare) from five countries in the West Asia and North Africa (WANA) region were studied using 36 simple sequence repeat (SSR) markers. The accessions were divided into subspecies/origin (S/O)‐groups and marker results were analysed in relation to genetic diversity and genetic structure. Wild barley from WANA was found to be highly diverse. The landraces from different countries of the Near East showed genetic diversity that was nearly as high as the wild barley from the same country. Further analyses showed that wild barley from Palestine/Israel represented the group with the highest diversity and the most complex structure. However, this group was distantly related to the cultivated barley in WANA, while the wild barley from the rest of the WANA region was closely related to the cultivated WANA barley. The high diversity and the close relationship to the wild barley make the WANA landraces an interesting genetic resource for both conservation and exploitation.     

Outcrossing rates of barley landraces from Syria

Diversity levels in populations of barley landraces may be influenced by varying levels of natural outcrossing caused by environmental conditions. Outcrossing was studied in 10 accessions of the barley landrace Arabi Aswad from different environments in Syria. Electrophoretic variation at two codominant isoenzymes (Est1 and Est2) in six seeds of 50 families per population were analysed and multilocus outcrossing rates calculated. Results were correlated with interpolated environmental conditions. Outcrossing was, on average, 1.7% and not significantly different from the outcrossing rate of wild barley. A significant increase of outcrossing was observed with increasing inter‐annual variation in rainfall and decreasing minimum temperatures of the coldest month.     

Radiation interception and radiation use efficiency of near-isogenic wheat lines with different height

Resistance to Pyrenophora teres Drechs. f. teres Smedeg., the net blotch pathogen, was studied in six 6-row Nordic spring barleys (Hordeum vulgare L.) in the field and in the greenhouse. The barley genotypes were: Arve, Agneta, Artturi, H6221, Pohto and WW7977. Disease progress was monitored in the field (1994 and 1995) in small artificially infected plots, sown at commercial seeding rates, and in infected hill plots (1994). Areas under the disease progress curves (AUDPC) and apparent infection rates (r) were calculated for the uppermost 3 or 4 leaves. Terminal severities (TS) were also recorded. Infection response of seedlings to a range of P. teres isolates was assessed in the greenhouse using a standard scale. In small plots in the field, Arve and Agneta were very susceptible to P. teres infection, as indicated by large values for AUDPC and TS. H6221 and WW7977 were highly resistant, while Artturi and Pohto were moderately resistant. In hill plots the situation was similar, except that Artturi and Pohto appeared less resistant than in the small plots. The relatively greater resistance of H6221 and WW7977 was reflected in seedling infection responses. According to the results of these experiments, H6221 and WW7977 possess adequate levels of quantitative resistance to P. teres to make them useful parents in future crossing programs aimed at improving net blotch resistance in Finnish spring barleys. This revised version was published online in July 2006 with corrections to the Cover Date.     

Resistance to powdery mildew in selections from Tunisian landraces of barley

Two-hundred and thirty-two accessions of barley landraces collected from Tunisia were screened for resistance to powdery mildew. A number of race-specific genes were detected using the detached leaf technique. Among the 232 accessions tested, 169 were susceptible to powdery mildew, 20 were resistant and 43 showed differential reactions to the three isolates of powdery mildew used. An attempt was made to determine the number of genes, the types of gene, the types of gene action and the gene loci in 20 resistant accessions. Three types of cross were made: (1) the accessions were crossed to the susceptible variety ‘Pallas’, (2) the accessions were crossed with ‘Pallas’ isolines, and (2) accessions with identical powdery mildew reaction patterns were intercrossed. Three isolates of Erysiphe graminis f. sp. hordei were used: Bzm-1, KM 18-75, R13C. A number of different resistance genes were detected among the 19 resistant accessions. Surprisingly, segregation indicating single genes only were detected with the isolates used. Some of these genes could be associated with loci already known. In 19 cases a dominant and in one a recessive mode of inheritance was detected. The recessive gene was not located at the mlo locus. This investigation represents the first systematic study of race-specific genes for powdery mildew resistance in Tunisian landraces. The newly identified sources of resistance may be used in many strategies of breeding for disease resistance.     

Net blotch resistance in wild species of Hordeum

Resistance to net blotch was evaluated in 175 Hordeum vulgare subsp. spontaneum (H. spontaneum) accessions and 149 accessions of thirteen species or subspecies of wild Hordeum. Most H. spontaneum accessions showed resistance to each of the four Pyrenophora teres f. teres (P. teres) isolates tested. However, H. spontaneum accessions showed different resistance reactions, depending upon their origin. In particular, some accessions from Afghanistan and Russia showed a high level of resistance, and accessions from Morocco were susceptible. Among the four P. teres isolates, the virulence spectra on the H. spontaneum accessions were more different between isolates from different countries than between those from the same country. Hordeum spontaneum accessions susceptible to the Canadian isolate WRS102, but resistant to the other three isolates were found in Iraq suggesting the geographical differentiation of resistance genes in H. spontaneum. All accessions of the other wild Hordeum species, especially some accessions of H. marinum subsp. gussoneanum, showed high levels of resistance. These resistance genes may be useful candidates for incorporation into cultivated barley. This revised version was published online in July 2006 with corrections to the Cover Date.     

New sources of resistance for stripe rust in barley

Eight spring barley accessions from the gene bank in Gatersleben, Germany, and 10 cultivars were tested for stripe rust resistance. Tests were performed at the seedling stage in the growth chamber and as adult plants in the field. All accessions and six cultivars were scored as resistant against race 24 under all test conditions, with very few plants as exceptions, while the susceptible control cultivars ‘Karat’ and ‘Certina’, and four other cultivars were attacked in all cases. Differences between accessions and between cultivars were detected after infection with isolates from ‘Trumpf’ and ‘Bigo’ (seedling tests only). Infection structures within seedling leaves without pustules and for the first time within leaves of adult plants from the field were analysed by fluorescence microscopy. With this method additional genetic Differences in the resistance reaction could be detected which could not to be seen in the resistance test. Crosses between the accessions and the susceptible cultivar ‘Karat’ led to segregating F₂ progenies. The percentage of resistant plants varied between the accessions. This also indicates a different genetic basis of resistance in the accessions. The infection structures observed by fluorescence microscopy stopped earlier in leaves of the two accessions HOR 8979 and HOR 8991 than in leaves of other accessions in all the tests. These accessions were the only ones with more than 50% resistant plants in all F₂ tests. In general, the accessions from the gene bank can be used as new resistance sources against stripe rust.     

The Effect of Seeding Rate and Nitrogen Fertilization on Barley Yield and Yield Components in a Cool Maritime Climate

Soil and climatic conditions in Newfoundland are on the margins of agricultural capability, and almost all feed grain is imported. The overall objective of this work was to develop guidelines for the production of barley in Newfoundland, with the goal of establishing modern cropping recommendations. We conducted a 4-year study near St. John's to examine the effect of seeding rate and topdress ammonium nitrate (N) fertilization rate on Chapais six-row barley ( Hordeum vulgare L.) yield components and grain yield. Increasing seeding rate from 200 to 380 seeds m^–2 did not alter grain yield in any year. Increasing topdress fertilization from 0 to 60 kg N ha^–1 increased spike density m^–2 at harvest, resulting in linear increases in grain yield in all years. Highest N rates had greatest lodging in two years. Based on our results, agronomic recommendations for eastern Newfoundland now include barley seeding rates of 250 ± 50 seeds m^–2, with topdress applications up to at least 30 kg N ha^–1.     

RFLP diversity within and between major groups of barley in Europe

Restriction fragment length polymorphism (RFLP) diversity has been determined and analyzed as expressed by 33 single‐ or low‐copy clone/ enzyme combinations at 32 loci distributed over all chromosomes of the barley genome within a sample of 223 European barley accessions comprised of pure line (single‐head progenies) genotypes. The accessions have been selected to include landraces and widely grown cultivars derived from crossbreeding during the 20th century in North‐, West‐ and Central European countries. Genetic diversity obtained from 83 alleles across all accessions is characterized by the diversity index H = 0.385. The diversity indices determined for landraces and cultivars were almost equal, with the difference between spring (H = 0.260) and winter (H = 0.415) barley approaching statistical significance, while comparisons of other groupings only revealed statistically insignificant trends. A more detailed analysis based on differences in allele frequency distributions at each locus (clone/enzyme combinations resp.) revealed very clear differences related to the existence, continuity and dynamics of changes in group‐specific RFLP profiles. With the majority (69%) of RFLP alleles at 23 out of 32 loci on all barley chromosomes involved, contributions from chromosomes 1H, 3H, 4H and 5H seem to be of special importance. Differences in the overall average of abundance indicate higher levels of genetic diversity within both groups of winter barley compared with both groups of spring barley, from which the most frequent alleles at 15 (2‐rowed spring barley) and 17 (6‐rowed spring barley) RFLP loci approach fixation. The results of this study are discussed in relation to the history of barley cultivation and barley breeding in Europe, and possible explanations for group‐specific differences in the RFLP profiles of landraces and cultivars as well as for the high levels of (nearly) fixed alleles of both subsets of spring barley, and with respect to progress in barley breeding that it has been possible to obtain within the rather narrow RFLP profiles.     

EcoTILLING for the identification of allelic variation in the powdery mildew resistance genes mlo and Mla of barley

In this investigation the successful implementation of a CEL I‐based mutation detection technique for the discovery and detection of DNA polymorphism in the genes mlo and Mla of barley is described. The technique is called EcoTILLING, which is a high‐throughput method to detect and discover new point mutations and small insertions/deletions in DNA. That the method not only reveals polymorphism between different alleles but can also be used as a powerful genetic marker is demonstrated. The genes mlo and Mla are involved in the defence of barley against the fungal pathogen powdery mildew. The powdery mildew resistance gene mlo is a single copy gene, whereas multiple alleles exist at the Mla locus. With EcoTILLING it was possible to identify point mutations and deletions in each of the 11 mlo mutants tested. For Mla 25 natural barley variants were tested, and although the identification was complex due to the presence of highly similar paralogues of Mla, most of the recently identified alleles from Hordeum vulgare ssp. spontaneum were identified. This method offers the possibility to combine different mlo alleles with different Mla alleles from wild barley to obtain cultivars with more durable resistance.     

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.     

Molecular mapping of greenbug (Schizaphis graminum) resistance gene Rsg1 in barley

The greenbug, Schizaphis graminum (Rondani) is an extremely damaging aphid pest of barley (Hordeum vulgare L.) particularly in the southern Great Plains of the USA. The simply inherited, dominant resistance gene Rsg1 is in all greenbug‐resistant US barley cultivars. In this study, we conducted molecular mapping of Rsg1 using an F_2:3 population derived from a cross between the greenbug‐resistant Post 90*4/R015 and susceptible CI2260 inbred lines. Segregation of host responses to greenbug biotype E infestation confirmed that a single dominant gene is responsible for greenbug resistance in Post 90*4/R015. Simple sequence repeat (SSR) markers evenly distributed along the seven barley chromosomes were employed for the construction of a framework genetic map. Linkage analysis placed the Rsg1 locus in the long arm of chromosome 3H (3HL) flanked by SSR markers Bmag0877 and GBM1420 that were 35 cM apart. Polymorphic single‐nucleotide polymorphism (SNP) markers in 3HL were identified from an Illumina GoldenGate SNP assay and used for targeted mapping to locate Rsg1 to an 8.4‐cM interval. Comparative analysis identified syntenic genomic regions in Brachypodium distachyon chromosome 2, in which 37 putative genes were annotated including a NB‐LRR‐type resistance gene homologue that may be a potential candidate gene for the Rsg1 locus of barley. Results from this study offer a starting point for fine mapping and cloning of this aphid resistance gene in barley.     

入侵植物印加孔雀草与本地植物青稞的竞争效应

为了探究入侵植物与本地植物之间的竞争关系,以入侵植物印加孔雀草和青稞为试验材料,采用添加系列试验法,在温室内设置了单种和不同比例混种方式进行控制试验。试验结果表明：（1）种间竞争降低了印加孔雀草的生物量、含水量,增加了其株高;降低了青稞的生物量、含水量和株高。（2）印加孔雀草的相对产量(RY_Tm)在0.78~0.98,差异不显著;青稞的相对产量(RY_Hv)在0.09~0.73,差异显著;混种对植物的影响为印加孔雀草小于青稞。（3）印加孔雀草和青稞的相对产量总和(RYT)在0.53~0.86,二者利用共同资源且有竞争关系。（4）印加孔雀草对青稞的竞争攻击力系数(A)在0.23~0.80,印加孔雀草竞争力强于青稞。研究表明：印加孔雀草和青稞混种时种间竞争>种内竞争,印加孔雀草的竞争力强于青稞,印加孔雀草影响青稞正常生长。