Embryo development in two barley genotypes after self pollination and pollination with Hordeum bulbosum L.

Summary Embryo development in vivo was compared in the two barley genotypes VK 16032 and Vogelsanger Gold after self pollination and after pollination with H. bulbosum. Embryo growth in VK 16032 after crossing with H. bulbosum showed a similar growth pattern to that of the self pollinated embryos, although the size increase was smaller. The hybrid embryos continued to grow until day 18 after pollination. The embryos from the cross between Vogelsanger Gold and H. bulbosum ceased to grow 8 days after pollination. Arrested embryo growth with subsequent abortion in Vogelsanger Gold was associated with a very early depletion and break down of the endosperm. Use of barley genotypes less sensitive to the genomic disturbances after crossing with H. bulbosum in combination with genotypes like Vogelsanger Gold is recommended in haploid breeding work.     

Pleiotropic effects on the ml-o mildew resistance gene in barley in different genetical backgrounds

Summary The effects on quantitative characters of the ml-o5 allele has been investigated by means of doubled haploid lines produced by the Hordeum bulbosum method. In one series of experiments the genetic background in each cross was found significantly to modify the negative pleiotropic effects. Doubled haploids were also produced from a F1 cross and tested in the field during two years, one with strong and one with weak mildew infections. In the first year only an insignificant reduction in grain yield was detected in the mlo lines, in the second it was significantly reduced with 5.9%. Thousand grain weights were, however, higher in the resistant lines. In both series of experiments the possibility to extract very high yielding mlo genotypes was demonstrated. The nature of the genetic background effect is discussed.     

Application of a set of 14 c-DNA probes from wheat to detect restriction fragment length polymorphism (RFLP) in barley

Summary A set of 14 probes from wheat cDNA clones was used to search for restriction fragment length polymorphism (RFLP) in six barley lines. The degree of polymorphism among the lines varied greatly between probes and between the various restriction enzymes. Two probes revealed a high degree of polymorphism in all probe/enzyme combinations. Seven of 14 probes did not reveal RFLP. The average level of polymorphism based on all 840 pairwise comparisons was 14.0%, which is higher than has been reported in wheat, but lower than in maize, rice, potato and lettuce. Most of the probes that detected RFLP correspond to sites on the long arms of wheat chromosomes.     

Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants

Summary Transgenic barley plants (Hordeum vulgare L. cv. Kymppi) were obtained by particle bombardment of various tissues. Immature embryos and microspore-derived cultures were bombarded with gold particles coated with plasmid DNA carrying the gene coding for neomycin phosphotransferase II (NPTII), together with plasmid DNA containing the gene for -glucuronidase (GUS).Bombarded immature embryos were grown to plants without selection and NPTII activity was screened in small plantlets. One plant proved to be transgenic (T₀). This chimeric plant passed the transferred nptII gene to its T₁ progeny. The presence of the nptII gene was demonstrated by the PCR technique and enzyme activity was analyzed by an NPTII gel assay. Four T₀ spikes and 15 T₁ offspring were transgenic. The integration and inheritance was confirmed by Southern blot hybridization. Transgenic T₂ and T₃ plants were produced by isolating embryos from green grains of transgenic T₁ and T₂ plants, respectively and growing them to plants. After selfing, the ratio of transgenic to non-transgenic T₂ offspring was shown to follow the rule of Mendelian inheritance. The general performance of transgenic plants was normal and no reduction in fertility was observed.Microspore-derived cultures were bombarded one and four weeks after microspore isolation. After bombardment, cultures were grown either with or without antibiotic selection (geneticin ^R or kanamycin). When cultures were grown without selection and regenerated plants were transferred to kanamycin selection in rooting phase, one out of a total of about 1500 plants survived. This plant both carried and expressed the transferred nptII gene. The integration was confirmed by Southern blot hybridization. This plant was not fertile.     

Inheritance and geographical distribution of phenol reaction-less varieties of barley

Summary The reaction of spikes and grains of barley to phenol was investigated using 8,849 cultivated and 349 wild accessions collected from the world. The pericarp and hull of the grain were stained dark brown by a 1% phenol solution and the reaction of awn was sharpest. Phenol reaction was controlled by a dominant gene, named Phr (phenol reaction) which was located on chromosome 2. All the wild strains of various Hordeum species showed a positive reaction to phenol indicating it was the prototype of the trait. Only 51 accessions of cultivated barley showed negative reaction to phenol. They were distributed along the so-called Silk-road and the type of variety was limited suggesting that it was a rather new mutation which occurred in the Middle East. Synteny of the chromosome region involving the phenol reaction gene in some gramineous plants was discussed.     

Studies for assessing the influence of hardening on cold tolerance of barley genotypes

Summary Frost tolerance of 30 barley (Hordeum vulgare L.) cultivars have been field evaluated in North Italy during the 1990/1991 winter season that was characterized by exceptionally low temperatures without snow cover. The results showed a significant correlation between cold injury and grain yield loss (r=0.61^**). Five cultivars chosen for their varying degree of frost tolerance were further evaluated using laboratory tests. Measurements of survival rate and membrane damage were used to assess the influence of hardening on frost resistance. The reliability of the tests is shown by the high correlation to the field data. For both the laboratory temperature regimes and field conditions, the tested cultivars showed the same order of classification. The effect of a rise in temperature at the end of the hardening treatment on frost tolerance is also reported. The laboratory tests here proposed can be integrated in a breeding programme for improving frost tolerance in barley.     

Experiments on anther culture in barley. Influence of culture methods on cell proliferation and organ differentiation

Summary Several experiments have been performed in order to induce cell proliferation and plant differentiation from pollen grains by anther culture in barley. Some modifications of the culture media, pretreatments and transfer of the anthers increased notably the frequency of cell division, but the low proportion of normal green plantlets differentiating from microspores and calluses remains the major obstacle preventing the practical use of anther culture in barley breeding.     

Inheritance of sensitivity to the insecticide diazinon in barley and the geographical distribution of sensitive varieties

Summary A total of 5,560 barley varieties collected from the world was tested for the sensitivity to an organophosphorous insecticide, diazinon. Among them, 708 (12.7%) varieties were sensitive to diazinon. The sensitive/insensitive reaction was clear when 1,000–2,000ppm diazinon solution was sprayed onto the young seedlings at low temperature. The sprayed leaves of the sensitive varieties wilted and failed to recover, while the succeeding leaves were normal and appeared to be unharmed although the vigor was reduced. Diazinon sensitivity was controlled by a dominant gene, named Diz, and it was located on chromosome 1 in an order of br-Diz-gl-5-n. The sensitive varieties were distributed mainly in the western part of the world, and none of the local varieties east of Nepal was sensitive to diazinon. Thus, Diz is a new and effective marker gene for studying the phylogeny of barley.     

Evaluation of methods for the assessment of malting quality in barley breeding

Summary Methods for assessing malting quality in barley breeding were evaluated for their precision, including variation between different batches, analysts and barley samples. The barley characters measured were grain moisture, ground grain moisture (two methods), grain weight, steep moisture, malt moisture (two methods), malt yield, malt nitrogen, malt soluble nitrogen, Kolbach index, malt diastatic power, malt -amylase, malt -glucanase, wort refractive index, hot water extract, extract yield, wort reducing sugars, wort total carbohydrates, wort -amino nitrogen and wort total nitrogen. The value of these measurements in barley breeding is discussed.     

Endosperm protein accumulation in wild and cultivated barley and their cross grown in spike culture

Summary The high protein wild relatives of cultivated cereals have proven difficult to utilize in plant breeding by direct selection for high grain protein percentage, and hence alternative selection criteria are needed. In this study, a spike culture method was used to measure differences in protein accumulation between wild and cultivated barley, and their cross, at different levels of nitrogen supply. Three genotypes, barley cultivar Hordeum vulgare L. cv. Clipper, a wild barley accession H. spontaneum Koch line 363, and a high protein F₅ line (38.4) derived from their cross, were grown from 8 to 27 days after flowering in in vitro spike culture. Nitrogen supply in the culture medium was either 0.4 g/l or 2.0 g/l of N supplied as NH₄NO₃. Spikes were harvested at approximately 3 day intervals during grain development, and salt soluble and hordein protein fractions were measured. Lines 363 and 38.4 differed from Clipper in having extremely high initial rates of protein accumulation, even at 0.4 g/l N. In high nitrogen conditions all three genotypes reached similar salt soluble plus hordein protein levels. Hordein-1 and hordein-2 fractions were measured separately; the percent of hordein-1 was higher in lines 363 and 38.4 than in Clipper at 0.4 g/l N. For all parameters measured, pot-grown spikes of matching age were harvested and were shown to be similar to the 0.4 g/l N treatment. The possible utilization of spike culture for identification of critical protein accumulation parameters is discussed, in relation to their possible utilization in breeding.     

Genotypic and environmental influence upon the nutritional composition of barley grain

Summary The grain yield and contents of the quantitatively predominant nutritional constituents of barley grain were determined in nine adapted spring barley varieties each grown at seven European locations with three or four replications.The largest variation in nutritional composition was due to different environmental conditions, but genotypic effects were also present. Interactions between genotype and environment were small. The average protein content at different locations varied from 8.1 to 14.7 per cent of the grain dry matter, and was not simply related to the amount of fertilizer-N applied.The nutritional composition of the grain was influenced by the grain yield level. The percentage of dietary fiber and protein decreased with increasing grain yield, but some varietal differences which were independent of the grain yield level could be established.The protein quality depended upon the protein level, as the protein contained more prolamin relatively to non-prolamin protein at high than at low protein levels. A difference between two varieties in the prolamin/non-prolamin ratio was consistent over a wide range of variation in protein content.     

Potential and limitations of isozymes for chromosomal localization of resistance genes against barley mild mosaic virus (BaMMV)

Summary To assess the possibilities offered by isozymes to locate resistance genes against barley mild mosaic virus (BaMMV), the isozyme patterns of 19 barley (Hordeum vulgare L.) genotypes carrying genes different from ym4 were determined. Of the 15 isozyme systems tested, only three were polymorphic, namely aconitate hydratase, esterase, phosphogluconate dehydrogenase, providing markers on four of the seven barley chromosomes. Studies of F₂ progenies derived from three crosses between resistant genotypes and susceptible varieties failed to reveal linkage between resistance genes and isozymes. Another goal of the experiment was to study the linkage relationships between ym4 and the esterase locus (Est1-Est2-Est4). Our estimates of the recombination rate between these two loci (3.41 and 8.32%) were in the range of those reported between these esterases and one of the resistance genes of the Chinese variety Mokusekko 3.     

Haploid barley from the intergeneric cross Hordeum vulgare x Psathyrostachys fragilis

Summary The intergeneric hybrid Hordeum vulgare x Psathyrostachys fragilis was fairly easily obtained. During each growing season the intermediate, perennial hybrid yielded haploid tillers of H. vulgare. Late in one season few, hybrid tillers headed. The morphology, cytology and enzymatic patterns of hybrid and haploid tillers were investigated.     

Chlorophyll fluorescence transient as a genetic marker of productivity in barley

Summary The photosynthetic characteristics of the penultimate leaf in barley genotypes (reciprocal F1 combinations and varieties) were studied at the flowering stage in order to analyze the relationship between these traits and the biological yield and the economic yield. The biological yield was correlated with the leaf area whereas the economic yield was more dependent on the green area duration of the leaf or the combination of the two factors, leaf area and green area duration. The net photosynthesis per unit leaf area, determined at anthesis, was not a factor of productivity. On the other hand, the chlorophyll fluorescence (P), which was negatively correlated with the leaf area (+0.5) and with the combination of leaf area and green area duration (+0.5), was found to be a good marker of the economic yield. In view of its high heritability, it may provide an effective selection criterion.     

The assessment of variation in two populations of Hordeum bulbosum L. for improving success rates in a doubled haploid barley programme

Summary Improvements in the success of doubled haploid production have been achieved partly by screening new accessions of Hordeum bulbosum. In order to assess levels of variation within stocks of this species, rates of embryo differentiation from the cross between H. vulgare × H. bulbosum were recorded for selections derived from two different stocks of H. bulbosum. There was little difference within stocks for this character despite variation in the morphology and banding patterns of two enzyme systems of one of the stocks. It is proposed that to obtain further increases in success rates a few selections from many accessions of H. bulbosum should be screened rather than many selections from a few stocks.     

Development of SCAR markers linked to a scald resistance gene derived from wild barley

The F₂ progeny of a third backcross(BC₃) line, BC line 240, derived from a Turkish accession of wild barley (Hordeum vulgare ssp. spontaneum),segregated for resistance to scald (Rhynchosporium secalis) in a manner indicating the presence of a single dominant resistance gene. Two SCAR marker slinked to this resistance were developed from AFLP markers. Screens of disomic and ditelosomic wheat-barley addition lines with the SCAR markers demonstrated that the scald resistance gene is located in the centromeric region of barley chromosome 3H,a region previously reported to contain a major scald resistance locus, Rrs1. Markers that flank the Rrs1 locus were used to screen the wild barley-derivedBC₃F₂ population. These markers also flank the wild barley-derived scald resistance, indicating that it maps to the same locus as Rrs1; it may beallelic, or a separate gene within a complex locus. However, BC line 240 does not respond to treatment with the Rhynchosporium secalis avirulence factorNIP1 in the same way as the Rrs1-carrying cultivar Atlas46. This suggests that the scald resistance gene derived from wild barley confers a different specificity of response to theRrs1 allele in Atlas46.In order to increase the durability of scald resistance in the field, we suggest that at least two scald resistances should be combined into barley cultivars before release. The scald resistance gene described here will be of value in the Australian environment, and the several markers linked to it will facilitate pyramiding. This revised version was published online in July 2006 with corrections to the Cover Date.     

The effect of anther orientation on microspore-derived plant production in barley (Hordeum vulgare L.)

Summary The effect of anther orientation on culture response has been examined in a range of barley cultivars of agronomic importance. Anther orientation did not significantly affect the percentage of anthers responding. However, orientation of the anthers in the up position with one lobe in contact with the medium significantly increased the number of green plants produced when compared to anthers cultured in the flat position. Green plantlet production was observed directly from embryoids i.e. without an intermediate callus phase. Cytological examination of the regenerants indicated that the majority of the plantlets possessed the normal barley karyotype. The implications of these results for Barley breeding are discussed.     

The influence of genotype and environment on seed and embryo development in barley (Hordeum vulgare L.) after crossing with Hordeum bulbosum L.

Summary Seed and embryo development was studied in crosses between H. bulbosum and the barley genotypes VK 16032, Vada and Vogelsanger Gold and subsequently the F₁ and F₂ progenies from VK 16032 x Vada and VK 16032 x Vogelsanger Gold. Both seed and embryo development are strongly influenced by the barley genotype. Favourable environmental conditions can promote seed and embryo development in genotypes with existing good characters. Dominant inheritance for good seed development and incomplete dominance for large embryos is evident. Linkage between small embryo size and winter habit exists in Vogelsanger Gold.     

Pleiotropic association of infection frequency and latent period of two barley cultivars partially resistant to barley leaf rust

Summary Cebada Capa, carrying four to six minor genes for a longer latent period (LP), was crossed to L94 and Vada, carrying no and five to six minor genes for a longer LP respectively. Of each of 68 F3-lines the infection frequency (IF) and the LP of ten just-heading plants were assessed. There appeared to be a strong association between IF and LP, whereby the relationship between IF and LP of both crosses could be described by a single linear regression equation. The data strongly suggest that the genes for increased LP pleiotropically decrease the IF. The possibility of a close linkage between genes for reduced IF and genes for increased LP, although unlikely, could not be excluded.     

Transgressive segregation in inter and intraspecific crosses of barley

Summary Six populations of F₂-derived lines of barley and their parents were evaluated for heading date, plant height, grain yield, bundle weight, and harvest index in a replicated experiment in the field. These data were used to estimate the minimum number of effective factor pairs segregating for each trait, the number of favorable factors contributed by each parent in a cross, and the frequencies and magnitudes of transgressive segregates.Heading date, plant height, and harvest index were controlled by three to four effective factor pairs, whereas grain yield and bundle weight were controlled by five or more.All three H. spontaneum strains used in our study contributed one or more useful genes for each of the traits, grain yield, heading date, plant height, bundle weight, and harvest index. Therefore, it seems that H. spontaneum can be a useful source of favorable genes for quantitative traits, especially for grain yield, which could be incorporated into barley varieties readily by backcrossing. Transgressive segregates for grain yield in the interspecific crosses may provide the basic materials for improving the productivity of cultivated barley varieties.Journal Paper No. J-9760 of the Iowa Agric. and Home Econ. Exp. Stn., Ames, IA 50011. Project 2227. Supported in part by a grant from the Science and Education Administration-Cooperative Research, United States Department of Agriculture.