The use of reversed-phase high performance liquid chromatography as an aid to the identification of European barley cultivars

Summary A reversed-phase high performance liquid chromatography (RP-HPLC) system was used to separate the storage proteins (hordeins) extracted from European barley cultivars. From a total of 38 barleys tested, 26 types of hordein patterns could be distinguished after RP-HPLC. This appears to be a marked improvement in resolution over that achieved in a similar survey of European barley cultivars using SDS polyacrylamide electrophoresis (32 hordein patterns resolved by SDS PAGE from a total of 160 spring and winter barleys tested).Different hordein patterns were resolved by RP-HPLC within each of two groups of barley previously classified by SDS PAGE as indistinguishable within groups (three distinct patterns identified in a total of five cultivars tested from group 1A and five patterns observed among eight cultivars from group 3B). Thus RP-HPLC achieves a higher resolution than undirectional electrophoresis and promises to be a valuable aid in the identification of European barley cultivars.     

Genetic mapping of the barley Rrs14 scald resistance gene with RFLP, isozyme and seed storage protein markers

The scald susceptible barley cultivar ‘Clipper’ and a third‐backcross (BC₃) line homozygous for the Rrs14 scald resistance gene that originally came from Hordeum vulgare ssp. spontaneum were grown in replicated field trials. The level of resistance that Rrs14 confers against field populations of the pathogen Rhynchosporium secalis, the causal agent of scald disease, was evaluated. The Rrs14 BC₃ line exhibited 80% and 88% less leaf damage than ‘Clipper’ in 1995 and 1996, respectively. Given this effectiveness of Rrs14, research was undertaken to identify a linked marker locus suitable for indirect selection of Rrs14. Based on linkage to a set of previously mapped loci, Rrs14 was positioned to barley chromosome 1H between the seed storage protein (hordein) loci Hor1 and Hor2, approximately 1.8 cM from the latter locus. The Hor2 locus is thus an ideal codominant molecular marker for Rrs14. The tight linkage between Rrs14 and Hor2 and the availability of alternative biochemical and molecular techniques for scoring Hor2 genotypes, permits simple indirect selection of Rrs14 in barley scald resistance breeding programmes.     

Geographical, altitude and agro-ecological differentiation of isozyme and hordein genotypes of landrace barleys from Ethiopia: implications to germplasm conservation

An analysis of the variability of genes encoding six isozyme systems (15 loci) and two storage proteins (2 loci) in landrace barley from Ethiopia is reported. The materials consisted of populations collected from sites as low as 1650 and as high as 3750 meters, covering a wide range of agro-ecological conditions and geographical areas. Of the 17 loci 7 were polymorphic and 10 monomorphic when the 95% criterion of polymorphism was applied. Despite the disproportionate monomorphic loci, polymorphism was detected in all populations when this criterion is used. The populations were found to possess fairly low mean number of alleles per locus (A = 1.5), low mean value of expected heterozygosity (H = 0.134) and a fairly high mean percentage of polymorphic loci (P = 35.3%). The mean F_ST= 0.474 for the populations is typical of inbreeding species. The result indicated that allelic richness is concentrated in altitude class 3 (2500–3000 m) followed by altitude class 1 (<2000 m). Altitude class 2 (2000–2500) holds an intermediate place though it is the highest in terms of expected heterozygosity (H = 0.245). Higher genetic diversity is concentrated in some geographical regions such as Shewa, Arsi, Bale compared to others (Welo, Gamu Gofa, Gojam). Genetic differentiation among the agro-ecological zones was more profound than both among the altitudes and among regions. Correlation analysis between phenotypic diversity (Shannon-Weaver diversity index) and expected heterozygosity (H) for isozyme/hordein loci revealed non significant associations except with respect to agro-ecological zones. In general, it was detected that sites in highland areas in central and northern regions may be more desirable for in situ conservation than sites in peripheral regions in terms of isozyme/hordein diversity and current rate of varietal replacement.     

Malting enzyme activities, grain protein variation and yield potentials in the displaced genetic resources of barley landraces of Finland

Thirty-two unimproved landraces of barley ( Hordeum vulgare L.) based on single plant selections in SE Finland were studied. Some of the lines still had ancient features, e.g., rachis brittleness in the tip of the spike. The lines showed 17 different and 16 unique hordein patterns and three lines showed more than one pattern, suggesting heterozygosity, and six different residual grain protein patterns. The three enzyme activities (-amylase, -amylase and -glucanase) of grains germinated aseptically for 120 h were determined. The average activity levels were high compared with a standard of five global barleys and with those determined previously in wild barley (H. vulgare ssp. spontaneum (Koch) Archers et Graebn.) grown in Finland. The ssp. spontaneum sample of 257 accessions showed significantly (P<0.001) less variation in -amylase and significantly (P<0.001) lower mean activity of all three enzymes. The high variation of these chemotypes indicates great potential variation of possible use by breeders has been lost by the disappearance and displacement of local barleys with commercial cultivars since 1950 in Finland before which barley cultivation and adaptation to the local environment had occurred over more than 3000 years. Selection for currently preferable plant characteristics in the descendants of the cross of HA52 (a landrace selection) × Adorra discriminated the hordein pattern of HA52 not being directly selected. The best landraces outyield the standard cultivars especially when there was no lodging. Top yield and small grains appeared to be associated characteristics under the environmental selection pressure, conflicting with the man-made regulations of the EU.     

The assessment of genetic variation in Spanish primitive cultivars of barley, Hordeum vulgare L., by a combination of isozymes and hordeins

Five isozyme and endosperm reserve protein systems were analysed using electrophoretic techniques in order to investigate the genetic diversity of 222 accessions of Spanish, local varieties of barley, Hordeum vulgare L., maintained at the Centro de Conservación de Recursos Fitogeneéticos of the I.N.I.A. (Alcalá de Henares, Spain). The esterase (EST) isozymes provided more information than did the other systems analysed, showing a total of 14 variable markers. The cathodic peroxidases (CPX) and acid phosphatases (ACPH) were also polymorphic. Malate dehydrogenase (MDH) and glutamate oxaloacetate transaminase (GOT) were monomorphic. The hordeins showed patterns of up to 15 bands, the majority of which were very useful in distinguishing genotypes. 17.2% of accessions showed a uniform genotype, 29.8% showed practically identical genotypes and 53% showed mixtures of different genotypes. It is noteworthy that the use of only two systems (EST and hordeins) and the analysis of only six loci (Est-1, Est-2, Est-4, Hor-1, Hor-2 and Hor-3) is sufficient to reveal the genetic diversity of the collection.