Improved frost tolerance and winter survival in winter barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) by <Emphasis Type="Italic">in vitro</Emphasis> selection of proline overaccumulating lines

Embryogenic calli derived from anther cultures of the two-rowed winter barley cultivar Igri were plated on solid L3 medium containing the proline analogue hydroxyproline (Hyp), 10–20 mmol l^–1. Exposure to Hyp caused severe degeneration of most of the calli. Hyp resistant calli, distinguishable by their lighter colour and higher growth rate, and control calli not exposed to Hyp were plated on L3 regeneration medium. From 22,500 anthers exposed to Hyp 46 Hyp resistant regenerates were obtained, which were transferred to soil. After cultivation for 5–10 weeks at normal growth conditions they were cold hardened at 2 C under short day conditions together with control regenerates. Frost tolerance assays with segments of fully grown leaves of unhardened and cold hardened plants revealed that Hyp resistant regenerants were significantly more frost tolerant than the control regenerants. Improved frost tolerance was found also in the progenies R₁ to R₉, and genotypic segregation in the R₁ generation in a 1:2:1 ratio was indicated. Increased proline content was observed in the R₂ generation and in subsequent generations and was significantly (P 0.001) correlated with increased frost tolerance in the Hyp lines. Comparative studies of R₉ progenies from homozygous R₂ plants with the wild type Igri under field conditions in winter at three locations in Europe as well as crossing experiments confirmed the heritable improvement of frost tolerance and winter survival, respectively, in the Hyp lines. The results support the hypothesis that proline accumulation in cold acclimated winter barley plants is causally related to the acquisition of frost tolerance. Moreover, the described biotechnological procedure may be applicable in breeding programs for improved winter hardiness and possibly also for other stress tolerances.     

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.     

Winter Hardiness, Frost Resistance and Vernalization Requirement of European Winter Oilseed Rape (Brassica napus var. oleifera) Cultivars within the Last 20 Years

Cultivars of European winter oilseed rape cultivated in the second half of the 1970s and in the mid-1990s were screened for their winter hardiness, frost resistance and vernalization requirement. A strong correlation between winter hardiness and frost resistance in both groups of rape has been noticed. Among oilseed rapes cultivated in the late 1970s, low erucic acid and particularly double zero cultivars were less winter hard than high erucic acid cultivars. Double zero cultivars were characterized by lower frost resistance and lower vernalization requirement. A significant correlation between vernalization requirement and both frost resistance and field survival has also been shown. Frost resistance of the 1990s (double zero) cultivars was higher than that of double low cultivars from the late 1970s. Their vernalization requirement was still small and did not correlate with either frost resistance or winter hardiness. It was concluded that reduction in the content of glucosinolates in the 1970s involved decrease in winter hardiness and vernalization requirement of cultivars. During the following 20 years winter hardiness of double low cultivars has been improved, but vernalization requirements have not changed. As a result no correlation between winter hardiness and vernalization requirement in contemporary canola cultivars has been observed.     

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.     

Effect of hardening on frost tolerance and fatty acid composition of leaves and stems of a set of faba bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.) genotypes

Frost tolerance is a main component of winter-hardiness and improving it would promote faba bean (Vicia faba L.) cropping in cool-temperate regions. In many species, leaf fatty acid composition was found to be related to frost tolerance. The objective of this study was to determine, in a representative sample of genotypes, the effect of hardening on leaf and stem (1) frost tolerance and (2) fatty acid composition, and to seek correlations between them. First leaf, second leaf and stem of 31 faba bean genotypes were analyzed after hardening and without hardening. High frost tolerance of known winter genotypes and several experimental lines was shown. Hardening had a significant, positive effect on frost tolerance of all three organs. Stems were on average more frost tolerant than leaves. Hardening induced significant changes in the fatty acid composition: oleic acid decreased significantly in leaves by 3.24% and in stems by 1.77%, whereas linolenic acid increased in leaves by 6.28% and in stems by 9.06%. In stems, correlations between frost tolerance and fatty acid composition were not significant. Correlation coefficients strongly indicated that non-hardened oleic acid content, changes in oleic acid and in linoleic plus linolenic acid content in leaves partly explained their frost tolerance; 0.347 (P < 0.1) < |r| < 0.543 (P < 0.01). The results corroborate the importance of using genetic differences in the fatty acid metabolism in breeding grain legumes for frost tolerance.     

Relationship between cold resistance,heading traits and ear primordia development of wheat cultivars

Summary For breeding early heading wheat cultivars with resistance to frost damage which are well adapted to dry areas of West Asia and North Africa, the relationships between winter hardiness, ear primordia development and heading traits, i.e. veernalization requirement, photoperiodic response and narrow-sense earliness, were assessed using a total of 30 genotypes of wheat (Triticum aestivum L.) grown in an experiment in Syria. The results of artificial freezing tests indicated that cultivars with good winter hardiness were to be found only in the winter wheat cultivars which required 50 or more days of vernalization treatment. These winter wheat cultivars did not initiate internode elongation without vernalization even at 95 days after planting. Thus their ear primordia were still underground and were protected from frost injury at this stage. Photoperiodic response and narrow-sense earliness were not associated with winter hardiness and earliness of internode elongation, but were related to the number of days to heading after planting. This indicated the possibility for breeding early heading cultivars with winter hardiness and tiller frost avoidance by combining high vernalization requirement, short narrow-sense earliness and neutral response to photoperiod.     

冬小麦品种抗霜冻力的影响因素分析

应用移动式人工霜箱,研究了冬小麦幼穗抗霜冻力随发育期的变化规律。结果表明,随发育期进程,幼穗抗霜冻力呈下降趋势,从药隔前期开始进入低温敏感期。利用室内人工霜箱,在药隔前期对21个品种进行了抗霜冻力鉴定,进而分析了不同品种、冬春性、越冬抗冻力以及叶片可溶性糖含量等对冬小麦抗霜冻力的影响。结果表明,除少数品种有一定抗霜冻力外,多数品种抗霜冻力无显著差异。品种抗霜冻力与其冬春性和越冬抗冻力均无显著相关性,与叶片可溶性糖含量之间亦无显著相关性。     

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.     

Effect of common bunt on the frost resistance and winter hardiness of wheat (Triticum aestivum L.) lines containing Bt genes

In order to determine the effects of bunt inoculation on frost resistance and winter hardiness in lines containing resistance genes, the bunt [Tilletia foetida (Wallroth) Liro, T. caries (DC.) Tulasne] susceptibility of wheat lines containing bunt resistance genesBt1 to Bt10 and the effect of the year on the degree of infection were studied over six years from 1991 to 1997 in an artificial inoculation nursery. Uninoculated and artificially inoculated wheat plants were tested for frost resistance in the phytotron in 1995 and in the field in boxes in three years from 1994/95 to 1996/97. The line withBt10 was very resistant, lines with Bt5, Bt6, Bt8 and Bt9 were resistant, the line with Bt4 was moderately resistant, those with Bt2 and Bt3 were moderately susceptible, the line with Bt1 was susceptible and the line with Bt7 was very susceptible to the local bunt population in Hungary. Bunt incidence also varied over years. The frost resistance of the Bt lines was generally lower after bunt inoculation than that of uninoculated plants. The increased frost kill in inoculated plants was not correlated with the extent of varietal susceptibility to bunt. Some lines with resistance, namely those with Bt5 (1.6% infection), Bt8 (0.6%) and Bt10 (0.0%), suffered significantly greater frost kill in the young plant stage as the result of bunt inoculation. By contrast, the Bt7line had excellent frost resistance and winter hardiness but suffered the greatest extent of bunt infection, whereas the Bt6 line had good frost resistance and good bunt resistance. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of alien 5R(5A) chromosome substitution on ear-emergence time and winter hardiness in wheat-rye substitution lines

Ear emergence time and response to vernalization were investigated in 12 alien substitution lines in which a pair of chromosomes 5A of recipient spring wheat cultivars was replaced by a pair of chromosomes 5R of Siberian spring rye ‘Onokhoiskaya’. The recipients were 12 spring cultivars of common wheat, each carrying different Vrn genes. Spring rye ‘Onokhoiskaya’ had the Sp1 (now called Vrn-R1) gene for spring growth habit located on chromosome 5R, but its expression was weaker. The Vrn-R1 gene had no effect on growth habit, ear emergence time and response to vernalization in wheat-rye substitution lines. Ears emerged significantly later in the 5R(5A) alien substitution lines than in the recipient wheat cultivars with the Vrn-A1/Vrn-B1/vrn-D1 or Vrn-A1/vrn-B1/Vrn-D1 genotypes. No difference in ear emergence time was found between most of the 5R(5A) alien substitution lines and the cultivars carrying the recessive vrn-A1 gene. The presence of the Vrn2a and Vrn2b alleles at the Vrn2 (now called Vrn-B1) locus located on wheat chromosome 5B was confirmed.The replacement of chromosome 5A by chromosome 5R in wheat cultivars ‘Rang’ and ‘Mironovskaya Krupnozernaya’, which carries the single dominant gene Vrn-A1, converted them to winter growth habit. In field studies near Novosibirsk the winter hardiness of 5R(5A) wheat–rye substitution lines of ‘Rang’ and ‘Mironovskaya Krupnozernaya’ was increased by 20–47% and 27–34%, respectively, over the recurrent parents.     

Genes for frost resistance in wheat

Wheat varieties differ in their responses to low temperatures. Geneticstudies on frost resistance in wheat are difficult because the effects arequantitative in nature and thus require precise genetic material andreproducible experimental conditions. The detailed diallel analyses indicatedthat the inheritance of frost resistance is polygenic and mostly additive.Nevertheless, studies using monosomic, ditelosomic and substitution lineshave identified specific chromosomes that carry genes responsible for frostresistance. In particular, the chromosomes 5A and 5D appear to carrymajor genes. Using molecular markers (RFLP, AFLP) and recombinantsubstitution lines it was shown that the Vrn-A1 (vernalization) and Fr1 (frost resistance) loci were located closely linked on the distal portionof the long arm of 5A, but recombination between them was found (cM = 2). The RFLP markers Xpsr426 and Xwg644 were tightlylinked to the Vrn-A1 locus. Loci Vrn-D1 and Fr2are located on the long arm of 5D. Fr2 and Vrn-D1 arehomoeologous to Fr1 and Vrn-A1. A physical map of theVrn-A1 and Fr1 genes was constructed on chromosome 5Ausing deletion lines. This cytogenetically based physical map could be usefulin further work on genome mapping and gene cloning.     

Location of a gene for frost resistance on chromosome 5A of wheat

Summary A gene for frost resistance on chromosome 5A of wheat was located using single chromosome recombinant lines from the cross between the substitution line Hobbit (Triticum spelta 5A) and Hobbit. In this sample of recombinant lines the locus for frost resistance, designated Fr1, is completely linked to the locus Vrn1 controlling vernalisation requirement. The results can be explained by a pleiotropic action of the Vrn1 locus or close genetic linkage between Vrn1 and Fr1. Further detailed study is necessary to resolve these alternative hypotheses.     

Combining frost tolerance,high grain yield and good pasta quality in durum wheat

Growing in Central Europe winter instead of spring durum wheat would substantially increase yield potential but is currently hampered by the lack of knowledge of frost tolerance present in elite material. The objectives of our survey were to (i) study the genetic variability and heritability of frost tolerance and its association with other important agronomic and quality traits in durum wheat, (ii) examine the potential to combine frost tolerance with high quality and high grain yield and (iii) investigate the consequences of the heritabilities and associations among traits on the optimum design of a multistage selection programme for winter durum wheat. We investigated 101 elite winter durum wheat lines and four commercial checks in field trials at four locations. Four agronomic as well as nine quality traits were recorded. In addition, frost tolerance was evaluated using a semi‐controlled test resulting in high‐quality phenotypic data. Genotypic variances (σ²_G) were significantly larger than zero for all traits, and heritabilities were moderate to high. Several elite durum wheat lines exhibited a frost tolerance comparable to that of two frost‐tolerant Triticum aestivum varieties. Frost tolerance was not negatively associated with other important agronomic and quality traits. The high quality of the phenotypic data for frost tolerance evaluated in a semi‐controlled test suggests that this is a cost‐efficient approach to consider frost tolerance at early stages of a multistage durum wheat breeding programme.     

不同类型冬小麦品种拔节后幼穗低温敏感期的鉴定

摘要：以百农矮抗58、豫麦49－198、豫农949和偃展4110这4个小麦品种为材料,利用人工气候室处理,研究了冬小麦拔节期幼穗抗霜冻力随幼穗分化进程的变化规律。在药隔期前后对4个品种进行了抗霜冻力鉴定,进而分析了不同品种、冬春性以及幼穗发育进程等对冬小麦抗霜冻力的影响。结果表明：4个品种中百农矮抗58抗冻能力较强,其他几个品种抗冻能力相对较弱。随幼穗发育进程的推进,幼穗抗霜冻力总体呈下降趋势, 从雌雄蕊末期发育进入药隔期,幼穗的抗霜冻力骤然下降,雌雄蕊分化期之前以及药隔期之后,抗霜力的下降趋势比较平缓,表明雌雄蕊至药隔期这个阶段是冬小麦幼穗分化的低温敏感期。品种的抗霜性研究适宜在该时期展开。     

Mapping of genes involved in glutathione, carbohydrate and COR14b cold induced protein accumulation during cold hardening in wheat

Using some of the chromosome substitution lines developed from thecrosses of the donor Cheyenne to Chinese Spring we showed that theaccumulation of water soluble carbohydrates during different stages ofhardening was time dependent. Moreover there was a significantcorrelation between the rate of carbohydrate accumulation and the frosttolerance. The expression and regulation of a wheat gene homologous tothe barley cold regulated cor14b gene was compared in frost sensitiveand frost tolerant wheat genotypes at different temperatures. Studies madewith chromosome substitution lines showed that the threshold inductiontemperature polymorphism of the cor14b wheat homologous genewas controlled by loci located on chromosome 5A of wheat, while cor14b gene was mapped, in Triticum monococcum, onto the longarm of chromosome 2A^m. Our study on the effect of cold hardeningon glutathione (GSH) metabolism showed that chromosome 5A of wheathas an influence on the GSH accumulation and on the ratio of reduced andoxidised glutathione as part of a complex regulatory function during coldhardening. In addition, the level of increase in GSH content duringhardening may indicate the degree of the frost tolerance of wheat.     

Genome-wide association study for frost tolerance in rapeseed/canola (Brassica napus) under simulating freezing conditions

Rapeseed/canola seedlings can be easily damaged by spring frost, which can rupture the cells and kill the plant. Genetic variations for frost tolerance have known to exist within rapeseed/canola gene pool. A genome-wide association study (GWAS) was conducted using 231 diverged rapeseed/canola germplasm to find the significant markers of the freezing tolerance traits. The genotypes were obtained from 21 countries and comprised of spring, winter and semi-winter growth types. The genotypes were evaluated in plant growth chamber under simulated freezing conditions. Highly significant genotypic variation was observed for the freezing tolerance. The best three freezing tolerant germplasms (Rubin, KSU-10, and AR91004) were winter type, while the four most freezing susceptible germplasms (Polo Canada, Prota, Drakkar, and BO-63) were all spring type. No geographical or growth habit type clusters were identified by structure analysis in this mixed population. One QTL was identified that was located on chromosome A02. Six freezing/abiotic stress tolerance genes have been identified in this study.     

Improved frost tolerance and winter hardiness in proline overaccumulating winter wheat mutants obtained by in vitro-selection is associated with increased carbohydrate,soluble protein and abscisic acid (ABA) levels

In previous studies in vitro-selection of proline overaccumulating lines of winter wheat (Triticum sativum L. cv. Jo 3063) with increased frost tolerance was reported. These traits were found to be genetically stable. In the present study the improvement of frost tolerance (winter hardiness) under field conditions is confirmed for F₇ progenies of the mutants. Moreover, the mutants accumulated higher levels of glucose and fructose, soluble protein and abscisic acid (ABA) in addition to proline than the wild type under cold hardening conditions in a growth chamber as well as under cold hardening field conditions. ABA and proline levels peaked when the temperature decreased, whereas carbohydrate levels increased more slowly at decreasing temperature. Soluble protein levels also increased during cold hardening, but in addition showed sharp declines during frost periods. Increased carbohydrate levels of the mutants were associated with lower osmotic potential values. The differences in carbohydrate, protein and ABA levels between the mutants and the wild type are probably due to pleiotropic effects of the mutation.     

Comparative mapping and its use for the genetic analysis of agronomic characters in wheat

Summary The advent of molecular marker systems has made it possible to develop comparative genetic maps of the genomes of related species in the Triticeae. These maps are being applied to locate and evaluate allelic and homoeoallelic variation for major genes and quantitative trait loci within wheat, and to establish the pleiotropic effects of genes. Additionally, the known locations of genes in related species can direct searches for homoeologous variation in wheat and thus facilitate the identification of new genes. Examples of such analyses include the validation of the effects of Vrn1 on chromosome 5A on flowering time in different crosses within wheat; the indication of pleiotropic effects for stress responses by the Fr1 locus on chromosome 5A; the detection of homoeologous variation for protein content on the homoeologous Group 5 chromosomes; and the detection of a new photoperiod response gene Ppd-H1 in barley from homoeology with Ppd2 of wheat.     

Breeding winter hardy grasses

Summary Perennial grasses are vital for Norwegian agricultural production. The nature and extent of winter damage on grasslands is highly dependent on climatic conditions, and determines both persistency and yield. Physical stresses such as frost and ice encasement predominate in coastal regions with an unstable winter climate, while biotic stresses such as low temperature fungi are more common in the inland regions. Development of hardening depends on plant adaptation and climatic conditions during autumn and winter. New winter-hardy cultivars should be bred for wide adaptation to winter stresses. The genetic background for the most important character, freezing tolerance, seems to be of polygenic nature with mainly additive gene action. Selection for increased freezing tolerance has been effective over generations in grasses, and in most grass species ample variation still exists to be exploited by breeding. However, in some species like perennial ryegrass, modern biotechnological methods should be used to improve freezing tolerance and winter hardiness.