Evaluation of a semi‐controlled test as a selection tool for frost tolerance in durum wheat (Triticum durum)

Most durum wheat (Triticum durum) varieties possess only low winter hardiness due to their frost susceptibility. In North America and Central Europe, durum wheat is therefore typically sown in spring to circumvent the local winter conditions. However, the yield potential of durum in these regions could be much better exploited if durum varieties with increased frost tolerance were available, which could be sown in autumn. A factor limiting breeding for increased frost tolerance is the variation in the occurrence of frost stress across years. The ‘Weihenstephaner Auswinterungsanlage’ is a semi‐controlled test that exposes the plants to all weather conditions. Snow coverage of the plants, serving as frost protection, is prevented by the movable glass lid of the semi‐controlled test. In this study, different scorings for frost tolerance based on this semi‐controlled test were evaluated and compared with frost tolerance data in the field. Our results illustrate the potential of the ‘Weihenstephaner Auswinterungsanlage’ as an indirect selection tool for frost tolerance in durum breeding programmes, especially when regular frost tolerance data from the field are not available.     

Association between frost tolerance and the alleles of high molecular weight glutenin subunits present in Polish winter wheats

Subunits of high molecular weight glutenins strongly influence wheat bread making quality and can be associated with important agronomic traits. Polish winter wheats show a significant quantitative dominance of the null allele over the coding alleles of the Glu-A1 locus. To identify the causes of such skewed distribution, 116 F₅ lines obtained from six cross combinations were analyzed for their HMW glutenin subunits and 11 agronomic characteristics, such as plant height and uniformity, leaf blotch and leaf rust resistance, grain yield per plot, number of grains per ear, grain yield per ear, 1000 kernel weight, frost tolerance, total protein content and the SDS-sedimentation value. The SDS-sedimentation value, resistance to leaf blotch and frost tolerance showed statistically significant associations with the status of the Glu-A1 locus. It appears that chromosome 1A with the null allele at Glu-A1 carries a closely linked locus responsible for frost tolerance. With early strong selection for winter hardiness, the null allele of Glu-A1 becomes fixed in advanced breeding materials despite its strong negative impact on the end use quality.     

Phenotypic evaluation of floral and flowering traits with relevance for hybrid breeding in wheat (Triticum aestivum L.)

Hybrid breeding is a promising approach to increase the yield potential in wheat (Triticum aestivum L.). The profitability of wheat hybrids highly depends on a cost‐efficient system for hybrid seed production for which an adequate outcrossing in the male pool is of utmost importance. Employing a set of 51 elite winter wheat lines, we developed and evaluated phenotyping methods for floral and flowering traits with relevance for improved cross‐pollination. We observed significant genotypic variances and high heritabilities for most traits, including important traits like pollen mass and anther extrusion. Our results suggest the utility of the developed phenotyping approaches for applied plant breeding and the potential of the traits to assist in the design of the male ideotype for increased cross‐fertilization.     

Potential of genomic selection in rapeseed (Brassica napus L.) breeding

Genomic selection employs genome‐wide marker data to predict genomic breeding values. In this study, a population consisting of 391 lines of elite winter oilseed rape derived from nine families was used to evaluate the prospects of genomic selection in rapeseed breeding. All lines have been phenotyped for six morphological, quality‐ and yield‐related traits and genotyped with genome‐wide SNP markers. We used ridge regression best linear unbiased prediction in combination with cross‐validation and obtained medium to high prediction accuracies for the studied traits. Our results illustrate that among‐family variance contributes to the prediction accuracy and can lead to an overestimation of the prospects of genomic selection within single segregating families. We also tested a scenario where estimation of effects was carried out without individuals from the family in which breeding values were predicted, which yielded lower but nevertheless attractive prediction accuracies. Taken together, our results suggest that genomic selection can be a valuable genomic approach for complex agronomic traits towards a knowledge‐based breeding in rapeseed.     

Effect of 3B, 5A and 3A QTL for Fusarium head blight resistance on agronomic and quality performance of Canadian winter wheat

The objectives of this study were to investigate (i) the correlations between Fusarium head blight (FHB) index, deoxynivalenol (DON) accumulation and percentage of Fusarium‐damaged kernels (FDK) with agronomic and quality traits and (ii) the effect associated with the presence of single QTLs for FHB resistance on agronomic and quality traits in winter wheat. The population was derived from the cross between ‘RCATL33' (FHB resistance derived from ‘Sumai 3’ and ‘Frontana’) and ‘RC Strategy’. Parental lines and recombinant inbred lines (RILs) were genotyped with SSR markers associated with the 3B, 5A and 3A QTLs. The population was planted in FHB‐inoculated nurseries and in agronomy trials. Lines in the 3B QTL class had the lowest FHB index, DON content and FDK level and did not have a significantly lower yield, thousand kernel weight or protein content compared with the lines grouped in other QTL classes (including no QTL class). Marker‐assisted selection of the 3B QTL for FHB resistance into high‐yielding FHB‐susceptible winter wheat is the recommended approach for the development of lines with increased FHB resistance without significant yield and quality penalties.     

The potential of genomic‐assisted breeding to improve Fusarium head blight resistance in winter durum wheat

Durum wheat is the most important tetraploid wheat mainly used for semolina and pasta production, but is notorious for its high susceptibility to Fusarium head blight (FHB). Our objectives were to identify and characterize quantitative trait loci (QTL) in winter durum and to evaluate the potential of genomic approaches for the improvement of FHB resistance. Here, we employed an international panel of 170 winter and 14 spring durum lines, phenotyped for Fusarium culmorum resistance at five environments. Heading date, plant height and mean FHB severity showed significant genotypic variation with high heritabilities and FHB resistance was negatively correlated with both heading date and plant height. The dwarfing gene Rht‐B1 significantly affected FHB resistance and the genome‐wide association scan identified eight additional QTL affecting FHB resistance, explaining between 1% and 14% of the genotypic variation. A genome‐wide prediction approach yielded only a slightly improved predictive ability compared to marker‐assisted selection based on the four strongest QTL. In conclusion, FHB resistance in durum wheat is a highly quantitative trait and in breeding programmes may best be tackled by classical high‐throughput recurrent phenotypic selection that can be assisted by genomic prediction if marker profiles are available.     

Line selection for exploiting durum wheat (T. turgidum L. var. durum) local landraces in modern variety development program

Summary Pure line selection for grain yield applied within four durum wheat local landraces (Triticum turgidum L. var durum) was studied in an effort to determine: (i) the agronomic performance of selected line mixtures (bulks) and their value as potential new cultivars, (ii) the effectiveness of pure line selection for grain yield within local landraces and (iii) the correlated selection response of yield components and kernel quality traits. Four local durum wheat landraces were used. Two of them, Mytilini-1 and Mytilini-2 are still cropped in some rural areas of the island Mytilini whereas the other two, Limnos and Mavragani are maintained in the Greek Gene Bank. Following their preliminary evaluation for agronomic performance under current growing conditions, 100 individual plants from each landrace were randomly selected. Selected plants (pure lines) were field evaluated head to row during 1997–98 and 1998–99 growing seasons. Based on grain yield performance, bidirectional selection, high vs low, was practised within each landrace. Thus the five higher yielding lines and their corresponding lower ones from each landrace were identified. Selected lines, with no further progeny testing, were pooled using a balanced seed composite to form the high yielding five line bulk and its respective low counterpart. The resulting 16 bulks (eight high and eight low) along with their four source landraces and two commercial varieties, Mexicalli and Simmeto, used as checks, entered field testing for agronomic performance in two and one location during 1999–00 and 2000–01 growing seasons respectively. Data for grain yield and yield components (1000 kernel weight, number of kernels, and productive tillers) along with kernel quality data (protein content and vitreous) were recorded. Data for grain yield indicated that the landraces studied seemed to be line mixtures, as expected, since they responded to bidirectional phenotypic pure line selection. The high yielding bulks were of interesting agronomic performance, equivalent to or significantly better than that of the commercial checks and are worth further testing as potential cultivars. Selected high bulks of Mytilini 2 landrace significantly outyielded both the source landrace and the commercial checks maintaining the high protein content of the source landrace, which was significantly higher than checks. This performance provides evidence that the within landraces selection of lines combining high grain yield with high protein content seems to be feasible. In conclusion, evidence is presented that genetic variability seems to be available within the landraces studied which could be worth using in breeding programs. Furthermore the simple pure line selection breeding effort undertaken seemed to be effective in developing line bulks with promising performance and worth further testing as potential cultivars.     

Correlated responses of fatty acid composition, grain quality, and agronomic traits to nine cycles of recurrent selection for increased oil content in oat

Increases in the groat-oil content of oat (Avena sativa L.) increase the energy value of the grain and improve the feasibility of extracting oat oil for use as a vegetable oil. Nine cycles of recurrent selection for greater groat-oil content conducted in a genetically broad-based oat population resulted in dramatic increases in groat-oil content. Our objectives were todetermine if selection for greater groat-oil content affected fatty acid composition, grain quality traits (test weight and seed weight), or agronomic traits (straw yield, biomass, harvest index, heading date, and height). We evaluated 100 random lines from the base (C0) population and each of the nine selection cycle populations in three environments in order to estimate means, genetic variances, heritabilities, and genotypic and phenotypic correlations of grain quality and agronomic traits. We also evaluated 20 random lines from each population to estimate changes in fatty acid contents. Oleate and stearate contents increased over cycles of selection, as did the ratio of unsaturated to saturated fatty acids. Palmitate, linoleate, and linolenate contents and all grain quality and agronomic traits except harvest index decreased over cycles of selection. There was no evidence for reduced genetic variance or heritability in C9 for any trait, but the genotypic and phenotypic correlations between agronomic traits and oil content fluctuated over cycles. Selection for increased groat-oil content improved oil quality but reduced grain quality and agronomic performance of the population. This revised version was published online in August 2006 with corrections to the Cover Date.     

Response of Wheat Varieties from Semi-arid Regions of Jordan to Salt Stress

This study was carried out to determine the effects of salinity levels (control, 6, 12 and 18 dS m^?1) on germination, seedling growth, some agronomic traits and proline accumulation in leaves of nine wheat varieties adapted to semi‐arid areas of Jordan. The tested wheat materials included eight durum wheat varieties (Haurani 27, Acsad 65, Om Rabbeeh, Sham 1, Safra Ma’an, Katma, Al‐Samra and F8) and one bread wheat variety (Diel Harthon). Final germination percentage, shoot and seminal root length, and all growth and yield parameters were significantly (P < 0.05) decreased by increasing salinity level. Proline content was significantly (P < 0.05) increased by increasing salinity. There were significant variety × salt interactions (P < 0.05) on final germination percentage, seminal root length, grain yield and yield‐related traits indicating that the varieties responded to salt differently. Sham 1 did not show any decrease in germination ability at the different salinity levels. Haurani 27, Acsad 65, Al‐Samra and Diel Harthon showed a nonsignificant reduction in germination potential at low and intermediate salt levels. Safra Ma’an and Al‐Samra showed the lowest reduction in seminal root length at low salt level and consequently exhibited the lowest stress susceptibility index ‘S’ values. Grain yield‐based stress susceptibility index ‘S’ indicated that Haurani 27, Acsad 65, Katma, Al‐Samra, F8 and Diel Jardoon were more salt tolerant than Om Rabbeeh, Sham 1 and Safra Ma’an. In conclusion, a similar salt tolerance was observed at different growth stages in Haurani 27, Acsad 65 and Al‐Samra. Consequently, these three varieties could be considered as salt tolerant and accordingly they are suitable for durum wheat improvement. Furthermore, Sham 1 had the highest ability to germinate at high salinity level indicating that it has a genetic potential for salt tolerance, at least at this stage of its life cycle.     

Development of drought‐tolerant breeding lines derived from Helianthus annuus × H. argophyllus interspecific crosses

Helianthus argophyllus is a wild species known as “silver sunflower”, which possesses several traits, including morphological traits that increase drought tolerance. Therefore, introgressions between chosen cultivated lines and two H. argophyllus accessions were made, and segregating generations were established. Important agronomic traits including single heading, high pollen fertility, silver canopy (indicating more cuticular wax), and content of cuticular waxes and oil were selected over six segregating generations. The resulting F₆ lines showed introgression of water saving traits, as they had lower excise leaf water loss, with comparable yield to standard checks. However, these F₆ lines were late maturing and showed poor flowering synchronization between the cultivated and introgressed lines. Introgressed line “D‐22” was particularly promising as a breeding line, with superior agronomic and drought resistance traits. This line had the potential to be used as an inbred parental line for introgression of drought resistance traits into elite sunflower germplasm. Combining ability analysis of the introgressed lines further showed their potential for heterosis breeding or to be used as parental lines in breeding programme.     

Direct and indirect selection for drought tolerance in alien tetraploid wheat x durum wheat crosses

After the evaluation of numerous accessions of primitive wheats for yield components and morphophysiological traits related to drought tolerance (e.g., maintenance of high relative water content, RWC; photochemical quenching of chlorophyll fluorescence, qQ; and chlorophyll loss, chl, under moisture stress conditions), several accessions belonging to three species (Triticum dicoccum, T. polonicum, and T. carthlicum) were crossed with the improved durum wheat varieties Cham 1 and Om Rabi 5. A direct selection (F₂ progeny) for yield and an indirect physiological trait were applied on interspecific T. durum x T. dicoccum, T. durum x T. polonicum, and T. durum x T. carthlicum populations. Divergent selection was applied to validate the possible use of morphophysiological traits (root parameters, RWC, photochemical quenching, proline content, and carbon isotope discrimination) in selection, and to evaluate the resulting effects on yield. Heritability and selection response of these traits has been evaluated, and the impact of divergent selection for morphological and agronomic characters was studied under field conditions. The divergent populations were evaluated under different environmental conditions in France, Syria, and Yemen. Selection for morphophysiological traits related to moisture stress, such as root parameters, RWC and carbon isotope discrimination was possible due to high h² values and effective, resulting in high genetic gains. However, the effect of selection for these traits on yield stability needs to be further studied. Furthermore, a modified bulk method (F₂ 'progeny method') was developed. Direct selection for grain yield per plant in F₂ was carried out and yield per line in F₃ was evaluated under contrasting environmental conditions in France, Syria, and Tunisia. Results revealed that some F₃ lines were higher yielding than the improved durum wheat varieties Cham 1 and Om Rabi 5 under both stressed (Aleppo) and favourable (Montpellier) environmental conditions. Lines were evaluated in preliminary yield trials at Montpellier (France), Aleppo (Syria), and Constantine (Algeria). Results indicated that the use of related species combined with the use of the modified bulk breeding method is promising not only for increasing durum wheat yield in drought prone environments, but also for improving durum wheat yield stability across contrasting environments. Results of both breeding strategies are presented, and the potential advantages of using related tetraploid species in durum wheat breeding for drought tolerance are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic improvement trends in agronomic performances and end-use quality characteristics among hard red winter wheat cultivars in Nebraska

Evaluation of wheat cultivars from different eras allows breeders to determine changes in agronomic and end-use quality characteristics associated with grain yield and end-use quality improvement over time. The objective of this research was to examine the trends in agronomic and end-use quality characteristics of hard red winter wheat cultivars grown in Nebraska. Thirty historically important and popular hard red winter wheat cultivars introduced or released between 1874 and 2000 were evaluated at Lincoln, Mead and North Platte, Nebraska in 2002 and 2003. An alpha lattice design with 15 incomplete blocks of two plots and three replications was used at all locations. Agronomic (days to flowering, plant height, spike length, culm length, grain yield and yield components, and grain volume weight) and end-use quality (flour yield, SDS-sedimentation value, flour protein content, and mixograph time and tolerance) traits were measured in each environment. Highly significant differences were observed among environments, genotypes and their interactions for most agronomic and end-use quality characteristics. Unlike modern cultivars, older cultivars were low yielding, and less responsive to favorable environments for grain yield and yield components. Semidwarf cultivars were more stable for plant height than traditional medium to tall cultivars. All cultivars had high grain volume weight since it is part of the grading system and highly selected for in cultivar release. Modern cultivars were less stable than older cultivars for SDS-sedimentation and mixing tolerance. However, the stability of older cultivars was attributed to their having weak mixing tolerance and reduced SDS-sedimentation values. The reduced protein content of modern cultivars was offset by increased functionality, as measured by mixograph and SDS sedimentation. In conclusion, breeders have tailored agronomic and end-use quality traits essential for hard red winter wheat production and marketing in Nebraska.     

Silage yield and quality traits in elite maize hybrids and their relationship to elemental concentrations in juvenile plants

European silage maize is cultivated for animal feed and biogas production. We evaluated 210 factorial crosses of elite dent and flint lines in multilocation trials for agronomic and quality traits together with biomass and shoot concentrations of 10 elements in juvenile plants. Significant genotypic variances, mainly due to general combining ability variance of the dent lines, and high heritabilities were observed for dry matter yield (DMY) and quality traits. DMY was not correlated with quality traits, but methane fermentation yield (MFY) and metabolizable energy content (MEC) showed significant correlations with starch and fibre content. Concentrations of elements N,P,S,K were positively correlated with each other but only in few cases correlated with DMY and quality traits. Parent lines with contrasting P concentrations differed in root morphology traits. Results support DMY as primary trait for selection of silage maize hybrids, but MFY is of negligible importance in breeding for biogas contrary to MEC for animal feed. Neither biomass nor elemental composition of juvenile plants were of predictive value for final DMY or quality traits.     

Marker-assisted characterization of frost tolerance in barley (Hordeum vulgare L.)

Five molecular markers associated to two frost tolerance QTLs ( Fr-H1 and Fr-H2 ) were tested both on nine Turkish accessions, classified by breeders as highly frost-tolerant, and on a previously described sample of 26 barleys, winter, facultative and spring. Accessions were characterized in terms of frost tolerance under both field conditions and artificial freezing test at −12°C. The Turkish lines resulted to be equal or superior to the most tolerant European genotypes tested, showing that they can be used to improve the frost tolerance of the EU barley germplasm. The marker Hv BM5A ( Vrn-H1 and Fr-H1 ) resulted to be the best predictor for assisted selection within this germplasm, because of its high correlation between allelic variation and phenotypic traits. Only Hv CBF4 of the three Hv CBF markers tested at Fr-H2 was associated to the trait, but at lower significance than HvBM5A . The PCR-based molecular marker of Vrn-H1 can thus be used in barley breeding not only for selection of facultative and winter types, but also for fast routine selection of frost tolerant genotypes.     

New durum wheat with Hessian fly resistance from Triticum araraticum and T. carthlicum in Morocco

Hessian fly is an important pest of wheat on the North American continent and the temperate Mediterranean drylands. Yield losses caused by this insect in Morocco are the heaviest in the Mediterranean region and are estimated to be 36% on average. Genetic resistance to Mediterranean Hessian fly biotypes has not been found in durum wheat, although large numbers of durum accessions were screened. Genes for resistance were found in common wheat; some of which are transferable to durum. However, there is a need to broaden the genetic base for resistance in durum wheat. The objective of this work was to introgress resistance from selected Triticum araraticum and T. carthlicum accessions using multiple backcross methodology. The experimental recipient durum wheat included numerous adapted and high‐yielding lines. Testing for Hessian fly resistance under controlled conditions and field yield data showed that this programme yields Hessian fly‐resistant durum lines with good yields and adaptation.     

开花后不同光周期条件下大豆农艺性状和品质性状的QTL分析

以开花期相近的181个大豆重组自交系(RIL)为材料,研究开花后不同光照长度对大豆主要农艺性状的影响,并在利用SSR标记构建大豆遗传图谱的基础上,分别在长日(16 h)和短日(12 h)条件下检测与主要农艺性状及其光周期敏感度(PS)相关的QTL。结果表明,开花后光照处理对大豆农艺性状和品质性状有较大影响,不同性状的光周期敏感度差异明显,株高＞主茎节数＞蛋白质含量、脂肪含量＞百粒重＞单株荚数＞蛋白质和脂肪总量。利用复合区间作图法检测到12个与株高、主茎节数、单株荚数、百粒重、蛋白质和脂肪总量等性状及各性状对开花后光周期处理的敏感度相关的QTL,分别定位于A1、A2、B1、B2、C1、D1a、F、L等8个连锁群上。其中,在短日条件下检测到4个QTL,可解释的遗传变异范围在11.37%~26.63%之间;在长日条件下检测到3个QTL,可解释的遗传变异范围在11.84%~27.85%之间;检测到5个与不同性状光周期敏感度有关的QTL,可解释相对应性状表型变异的范围在6.15%~21.44%之间。针对同一性状,未检测到在长日和短日条件下均起作用的主效QTL, 说明开花后光周期对大豆产量和品质性状相关基因的表达有较大影响。     

Effect of genotypic,meteorological and agronomic factors on the gluten index of winter durum wheat

The determination of the gluten index is a widely used method for analysing the gluten strength of bread wheat and spring durum wheat genotypes. The present work was carried out to study the effect of the genotype, meteorological factors (temperature, precipitation and number of days with T_max ≥ 30 °C) and agronomic treatments (N fertilisation and plant protection) on the gluten index of winter durum wheat varieties and breeding lines. The results indicated that the gluten index had little dependence on the environment, being determined to the greatest extent by the genotype. Compared with varieties having weak gluten, those with a strong gluten matrix responded less sensitively to changes in environmental conditions. Among the meteorological factors, high temperature at the end of the grain-filling period caused the greatest reduction in the mean gluten index of three varieties (R ² = 0.462), while the fertiliser was found to be a significant factor affecting the gluten strength of winter durum wheat varieties. Using selection based on the gluten index, the gluten strength of winter durum wheat lines can be improved sufficiently to make them competitive with high quality spring varieties.     

Genetic variation in drought tolerance at seedling stage and grain yield in low rainfall environments in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Genetic architecture of seedling drought tolerance is complex and needs to be better understood. To address this challenge, we developed a protocol to identify the most promising drought-tolerant genotypes at the seedling stage in winter wheat. A population of 146 recombinant inbred lines (F₉) derived from a cross between wheat cultivars, ‘Harry’ (seedling drought tolerant) and ‘Wesley’ (seedling drought susceptible) were used in this study. All genotypes were sown in three replications in a randomized complete block design under controlled conditions in a greenhouse. Seven traits were scored and grouped into tolerance traits; days to wilting, leaf wilting, and stay green and survival traits; days to regrowth, regrowth, drought survival rate, and recovery after irrigation. Three selection indices were calculated (1) tolerance index, (2) survival index, and (3) drought tolerance index (DTI). The same set of genotypes were also tested for grain yield in two low rainfall environments for two seasons. High genetic variation was found among all genotypes for all seedling traits scored in this study. Correlations between tolerance and survival traits were weak or did not exist. Heritability estimates ranged from 0.53 to 0.88. DTI had significant phenotypic and genotypic correlations with all seedling traits. Genotypes were identified with a high drought tolerance at the seedling stage combined with high grain yield in low rainfall. Breeding for tolerance and survival traits should be taken into account for improving winter wheat drought tolerance at seedling stage. The selected genotypes can be used for to further improve drought tolerance in high yielding wheat for Nebraska.     

Accuracy of within‐ and among‐family genomic prediction in triticale

Genomic prediction has emerged as a powerful genomic tool to assist breeding of complex traits. In this study, we employed a population of 647 triticale doubled haploid lines derived from four families to assess the potential of this approach for triticale breeding. All lines were phenotyped for grain yield, thousand‐kernel weight, biomass yield, plant height, frost tolerance and Fusarium head blight resistance. The obtained prediction accuracies were moderate to high and consisted to varying degrees of within‐ and among‐family variance, in line with the different degrees of phenotypic differences between family means. The prediction accuracy within individual families also varied with the genetic complexity of the traits and was generally highest based on effect estimation with lines from the respective family, whereas the prediction accuracy decreased with decreasing relatedness among the families. Taken together, our results illustrate the potential of genomic prediction to increase selection gain in triticale breeding, but the composition of the training set is of utmost importance, and consequently, the implementation of this approach in applied breeding programmes is not straightforward.     

Heritable improvement of frost tolerance in winter wheat by in vitro-selection of hydroxyproline-resistant proline overproducing mutants

In a previous study (Dörffling et al., J. Plant Physiol. 142, 222–225, 1993) in vitro-selection and regeneration of hydroxyproline (Hyp)-resistant lines of winter wheat (Triticum aestivum cv. Jo 3063) with increased frost tolerance and increased proline contents was reported. In this study the heritability of these traits was investigated. The F ₁progenies derived from the pollination of regenerated plants with pollen from wild type plants developed higher mean frost tolerance (lower LT ₅₀values) and higher proline levels compared with in vitro-controls and wild type plants. In the F ₂generation, which was obtained by self-pollination, segregation of the Hyp-lines in a 3:1 phenotypic ratio with regard to the traits frost tolerance and proline content was observed. Subsequent selection of extremes in the F ₂yielded homozygous plants in the F ₃generation. Lower LT ₅₀values and increased proline contents in those F ₃plants were significantly correlated. Furthermore, improved frost tolerance of one of the selected mutants was also observed in studies with whole plants in the F ₄generation. These results, which are similar to results of parallel studies on winter barley, provide strong evidence for the heritability of the traits 'increased frost tolerance' and 'increased proline content' obtained by this in vitro-selection procedure. The mutation seems to be due to a single incompletely dominant gene. A comparison of frost tolerance values from F ₃Hyp-progenies with those of seven standard varieties differing in frost tolerance indicates a considerable improvement of frost tolerance by the described in vitro-selection method.