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.     

Comparisons between doubled haploid lines produced by anther culture,the Hordeum bulbosum-method and lines produced by single seed descent in barley crosses

Summary Possible negative effects of tissue culture on qualitative and quantitative characters have been investigated in three crosses of spring barley, using doubled haploid lines produced by anther culture (AC, with maltose as a carbon source) or the Hordeum bulbosum-method (HB). In one cross inbred lines produced by single seed descent (SSD) have also been included. Quantitative characters were investigated in a 2-year field experiment at one location. The results show that although the methods in the majority of cases gave similar results, the genotypic arrays produced were not identical. Different markers deviated in the various cross/method combinations. The ranking of methods as well as the frequencies of lines transgressive for grain yield differed between crosses. Thus no consistently negative impact of anther culture in barley has been found, and, although not identical, the various methods may be considered equivalent.Abbreviations AC Anther Culture method - SSD Single Seed Descent method - HB Hordeum bulbosum method     

Plant regeneration from embryo-callus culture in barley

Summary Plants were regenerated from callus cultures initiated from immature embryos of barley, Hordeum vulgare L. Immature embryos from seven diverse genotypes were cultured on modified Murashige and Skoog (MS) medium supplemented with 1.5 mg 2,4-D and 6.5 mg IAA/l. Of the 249 embryos cultured, 30% initiated callus within 8 days. Subculture of callus for 80 to 100 days on half-MS medium supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l zeatin resulted in organogenesis. Culture of organogenic calli for 30 days on half-MS medium without growth regulators produced plants which originated mostly via multiple shoot formation. Callusing response of the tested genotypes ranged from zero to 44%; however, only 23% of the calli were regenerative. Regenerated plants included variants for chlorophyll deficiency, plant height, stem thickness, spike shape, pollen fertility, seed set and ploidy.     

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.     

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.     

Selection environment and environmental sensitivity in barley

Summary Groups of 10 barley genotypes were selected for high grain yield under either high yielding (two groups) or low yielding conditions (two groups). The genotypes had a similar average grain yield across a wide range of yielding conditions, but differed in their linear response over environments (environmental sensitivity). The genotypes selected for high grain yield under low yielding conditions were less sensitive to changing environments than genotypes selected for high grain yield under high yielding conditions. The higher stability of genotypes selected under low yielding conditions was shown by both the linear regression analysis and the comparison of coefficients of variation. The use of a safety-first index showed that the probability of a crop failure of genotypes selected for high grain yield under high yielding conditions was between 1.8 and 2.7 times higher than for genotypes selected for high grain yield under low yielding conditions. The development of new cultivars for areas where a large proportion of the crop is grown by subsistence farmers should therefore be based on selection under low yielding conditions.     

Race differentiation and search for sources of resistance to Rhynchosporium secalis in barley in Italy

Summary Barley in Italy has recently been seriously affected by Rhynchosporium secalis. The pathogenic variation of the fungus was studied and 17 races were differentiated on 13 barley cultivars carrying most of the currently known genes for resistance. RC 1, the most virulent and most frequent race, was virulent on 10 out of the 13 differentials and the remaining races proved to be less virulent variants of RC 1. Atlas (C.I. 4118), Atlas 46 (C.I. 7323) and Osiris (C.I. 1622) were the only three differentials resistant to all the analyzed single-spore isolates.Differential cultivars previously assumed to have identical resistance factors did not react in the same way to all the Italian races, thereby revealing either undisclosed differences in the genes described or the presence of additional unidentified ones.Our findings were compared with previous data about virulence of scald populations from different countries, on the basis of tests with common differentials: fundamental differences were found between the Italian population and those of other countries with regard to virulence patterns.The susceptible reactions to race RC 1 of most barley cultivars grown in Italy indicate the urgent need for resistance genes to be incorporated in the cultivated material. Seventy-one barley accessions, known as sources of resistance in different parts of the world, were screened for their behaviour to races RC 1 and RC 13. Twenty-two appeared resistant to both of them.     

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.     

Simultaneous selection of segregating two-row winter barley material in England and Italy

Summary Independent but simultaneous selection by two breeding teams of the same segregating two-row winter barley material at Cambridge, England and Fiorenzuola, Italy was carried out over the years 1978–1982 with the objective of comparing the individual selection of the two breeding teams and assessing the adaptation of the surviving selections. Visual selection in the generations F₃ to F₅ did not result in any clear cut compartmentalisation of the material and both teams were as likely to select from families previously selected only by the other team as from families they had themselves previously selected. Lines with specific adaptation to each site as well as lines well adapted to both sites were identified from yield trials carried out in F₅ and F₆. The former showed only a very small yield advantage over lines with general adaptation to both sites.     

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.     

The accumulation of glycinebetaine during cold acclimation in early and late cultivars of barley

Summary Barley plants are able to accumulate glycinebetaine (betaine) at high levels in their leaves in response not only to water and salt stress but also to cold stress. Such accumulation of betaine during acclimation to cold is associated to some extent with freezing tolerance in leaves of barley plants, as previously demonstrated with near-isogenic lines that differed only in a single gene for the spring type of growth habit (Plant, Cell and Enyironment 17: 89–95, 1994). We now present evidence that the levels of betaine accumulated during cold acclimation might be associated with the earliness or lateness of the maturity of cultivars, namely, that late cultivars accumulate more betaine than early cultivars. Moreover, the grade of the vermalization requirement of the cultivars seemed unlikely to be associated with the level of betaine acumulated during cold acclimation. However, the trait that controlled accumulation of betaine during cold acclimation was not linked with the earliness or lateness of the maturity of cultivars. The higher levels of betaine in the late cultivars might have resulted from co-selection for lateness of maturity and freezing tolerance, which is generally a requirement in the areas of Japan where such late cultivars were originally cultivated.     

Aluminum tolerance in barley: Methods for screening and genetic analysis

Aluminum (Al) tolerance in roots of two cultivars of barley was studied using hematoxylin staining and root re-growth procedures. This study was performed in two F₂ segregating populations originated from crosses between the tolerant FM-404 and sensitive Harrington cultivars. The F₂ progeny analysed with hematoxylin staining revealed a segregation ratio of 3 tolerant: 1 sensitive, showing that the Al tolerance is controlled by a single gene with complete dominance for tolerance. The root re-growth measures do not confirm the 3:1 ratio. This last result can be explained due to the occurrence of genes that affect root growth rate or to the difficulties found in the evaluation of root re-growth. Barley has a complex root system, which makes it difficult to measure root re-growth after an extended period in nutrient solution. Due to the simplicity, reliability and better precision, the hematoxylin staining is the best procedure to determine the Al tolerance and its inheritance in barley. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

The grain yield of uniculm barley (Hordeum vulgare L.) in two contrasting environments

Summary Four six-rowed uniculm lines of spring barley were grown in two yield trials at different sowing densities. One trial was grown on light sandy soil in 1990. The second trial was grown on clay soil in 1991 and included the two-rowed, tillering cultivar Golf. In the trial on sandy soil, the grain yield of the uniculm lines approached that of Golf grown in an adjacent trial, but in 1991 when the growing conditions were more favourable, Golf yielded significantly more than the uniculm lines. Uniculm lines apparently perform relatively better under marginal growing conditions than in high yielding environments.The uniculm lines do not conform to the ideotype proposed for wheat by Donald (1968) but the results show that a drastic change in plant type need not to imply a large drop in grain yield.     

Cytogenetic variation in somatic tissue cultures and regenerated plants of barley (Hordeum vulgare L.)

Summary Chromosome number of morphogenic and non-morphogenic calli and regenerated plants of barley were determined. Cultures were obtained from two kinds of explants, immature embryos and seedling leaves from three cultivars, Ingrid, Dissa and Golden Promise. Callus chromosome analyses were carried out during a 12 month period in a medium containing 2 mg/l of 2,4-D. Diploid cells were predominant in all cases; although in leaf-derived cultures, retraploid cells (2n=4x=28) showed a tendency to increase as time in culture increased and after more than six months in culture, diploid cells decreased to percentages of almost 70%. Aneuploid cells were generally infrequent in all cases. The source of explant has been more important than the genotype (cultivar) and the type of callus (morphogenic vs. non-morphogenic) in the chromosomal stability of cultures as time increases. From short term cultures, only 1.85% of the regenerated plants were tetraploid, the remaining were diploids. The ability of morphogenic calli to regenerate plants decreased before any significant reduction of diploid cells were observed.     

An alternative approach to breeding for high diastatic power in barley

Summary Several lines from the cross Akka x Feebar were observed to have -amylase activity considerably in excess of either parent. It is suggested that, from crosses between two-rowed and six-rowed varieties, two-rowed genotypes with enhanced levels of grain nitrogen and -amylase activity may be obtained. These can be successfully exploited in a breeding programme to produce barley varities with high diastic power.     

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.     

Grain yield and its components in spring barley under row and hill plot conditions

Twenty homozygous barley lines were grown in row and hill plots at two locations in Ontario in 1973. The hill plots were of two spacings –30×30 cm and 45×45 cm between hills. Within each spacing, three seeding rates were used i.e. 15, 25 and 35 seeds per hill. Grain yield and three of its components, namely; number of seeds per spike (NSS), number of spikes per plot (NSP) and seed weight (SW) were studied. It was concluded that both NSS and SW could be evaluated in hill plots. NSS was found to be an important component of grain yield and its relationship with grain yield was unaffected by plot types and locations. SW also remained unaffected by plot types and seeding rates but as a component of grain yield it was the least important of the three components measured. The correlation between SW and grain yield could be changed by locations. NSP was as important as NSS as a component of grain yield, and was highly correlated with grain yield. Unlike NSS, NSP could not be efficiently evaluated in hill plots since its correlation between row and hill plots was generally lower.     

Exploitable genetic variation in a composite bulk population of barley

Summary Advanced agricultures are characterized by a great increase in the adoptation of uniform crop cultivars associated with a drastic reduction in locally adapted variation, the plant breeders should also examine means of creating and conserving genetic resources. Composite cross breeding is a technique that creates and preserves genetic variation in an exploitable form. The merits and problems of this method were discussed. using data obtained from barley Composite Cross XXI.     

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.