WCS120 protein family and proteins soluble upon boiling in cold-acclimated winter wheat

The amount of proteins soluble upon boiling (especially WCS120 proteins) and the ability to develop frost tolerance (FT) after cold acclimation was studied in two frost-tolerant winter wheat cultivars, Mironovskaya 808 and Bezostaya 1. Protein gel blot analysis, mass spectrometry (MS) and image analysis of two-dimensional gel electrophoresis (2-DE) gels were used to identify and/or quantify the differences in protein patterns before (non-acclimated, NA) and after 3 weeks of cold acclimation (CA) of the wheats, when FT increased from -4 degrees C (lethal temperature (LT(50)), for both cultivars) to -18.6 degrees C in Bezostaya 1 and -20.8 degrees C in Mironovskaya 808. Only WCS120 protein was visible in NA leaves while all five WCS120 proteins were induced in the CA leaves. Mironovskaya 808 had higher accumulation of three members of WCS120 proteins (WCS120, WCS66 and WCS40) than Bezostaya 1. MS analysis of total sample of proteins soluble upon boiling showed seven COR proteins in the CA samples and only three COR proteins in the NA samples of cultivar Mironovskaya 808 (MIR). In conclusion, the level of the accumulation of WCS120, WCS66 and WCS40 distinguished our two frost-tolerant winter wheat cultivars. Moreover, the differences of CA and NA samples of the MIR were shown by liquid chromatography (LC)-tandem mass spectrometry (MS/MS).     

Frost tolerance in winter wheat cultivars: different effects of chromosome 5A and association with microsatellite alleles

Frost tolerance of ten Bulgarian winter wheat (Triticum aestivum L.) cultivars (Milena, Pobeda, Sadovo-1, Enola, Kristal, Laska, Svilena, Russalka, No301 and Lozen) and five foreign cultivars (Mironovskaya 808, Bezostaya-1, Rannaya-12, Skorospelka-35 and Chinese Spring) was studied in two experimental seasons following natural cold acclimation and in one experiment carried out in controlled acclimation conditions. Considerable intercultivar variability in plant survival was observed after freezing at ?21 °C following sufficient cold acclimation, or at ?18 °C following insufficient or controlled acclimation. In seven cultivars, the effects of chromosome 5A on frost tolerance were investigated in their F₂ hybrids with chromosome 5A monosomic lines of cultivars with high, intermediate and low frost tolerance. The effects of chromosome 5A depended on the stress severity and the genetic background of the hybrids and varied even in cultivars of similar frost tolerance and vernalization requirements. Effects of other chromosomes besides 5A on frost tolerance were assumed. The analysis of six microsatellite loci located in the interval from centromere to Vrn-1 on of chromosomes 5AL, 5BL and 5DL showed that the major loci determining frost tolerance in Bulgarian winter wheats were Fr-A2 on chromosome 5AL, and, to a lesser extent, Fr-B1 on chromosome 5BL. A strong association of the 176 bp allele at locus wmc327 tightly linked to Fr-A2 with the elevated frost tolerance of cvs. Milena, Pobeda, Sadovo-1, Mironovskaya-808 and Bezostaya-1 was revealed. Relatively weaker association between frost tolerance and the presence of the 172 bp allele at locus Xgwm639 tightly linked to Fr-B1 was also observed.     

Beginning and duration of expression of photoperiodic response genes in winter common wheat

The influence of various Ppd genes on the beginning and duration of photoperiodic responce has been investigated in near isogenic lines of winter bread wheat Mironovskaya 808. During ontogenesis the photoperiodic responce is ascertained from the middle 2nd stage of ontogenesis according to Kuperman (usually a week later after vernalization completion in winter genotypes) and it is completed to the late 5th stage (2-3 weeks before heading). Different Ppd alleles do not affect the photoreaction intensity however they have an influence on its duration through the rate of development. Ppd-A1a and Ppd-B1a genes manifest shorter duration of expression when compared to the recessive alleles in the initial Mironovskaya 808 cultivar. Effect of the Ppd-B1a gene is stronger and of the Ppd-A1a is weaker.     

New data on the distribution of hybrid necrosis genes in winter bread wheat (Triticum aestivum L.) cultivars

Hybrid necrosis genotypes have been identified in 125 Russian cultivars of winter bread wheat. More than half of them (56%) carry the Ne2 gene (genotype ne1ne1Ne2Ne2); others are free of necrosis genes (genotype ne1ne1ne2ne2). The possible causes of the increase in the Ne2 allele frequency and the loss of the Ne1Ne1ne2ne2 genotype in modem Russian cultivars of winter wheat are discussed. The principal component method has been used to compare the structures of the genetic diversity of cultivars differing in the hybrid necrosis genotype. It has been found that the Ne2 allele in winter wheat cultivars from northern Russia has originated from the cultivar Mironovskaya 808, whereas the cultivar Bezostaya 1 is not a source of this gene. In cultivars from southern Russia, the presence of the Ne2 allele is also mainly accounted for by the use of Mironovskaya 808 wheat in their breeding. The recessive genotype is explained by the presence of descendants of the cultivar Odesskaya 16 in the pedigrees of southern Russian winter wheats. The genetic relationship of cultivars with identical and different necrosis genotypes has been analyzed in nine regions of the Russian Federation.     

Chromosomal location of genes for vernalization duration (Vrd) in winter bread wheat

In this study, we carried out monosomic analysis of two isogenic lines of cultivar Mironovskaya 808, monogenically dominant for genes Vrd1 and Vrd2 reducing the cultivar vernalization requirement duration from 50 to respectively 35 and 45 days, as well as analysis of substitution lines Cappelle Desperez/Bezostaya 1. Gene Vrdl is localized on chromosome 4A; Vrd2, on chromosome 5D. The third gene, which is not allelic to the former two, is present in cultivar Cappelle Desperez on either the IA, 6A, or 4B chromosome.     

The relationship between vernalization requirement and frost tolerance in substitution lines of wheat

Two sets of wheat (Triticum aestivum L.) substitution lines for the homoeologous group 5 chromosomes, 5A, 5B and 5D, carrying vernalization genes (Vrn-A1, Vrn-B1, Vrn-D1) were used to study the relationship between vernalization requirement and winter survival, with respect to the induction and maintenance of frost tolerance. Substitution lines carrying dominant Vrn loci substituted from the spring cultivars Zlatka (5A), Chinese Spring (5D) and the alternative cultivar eská Pesívka (5B) into three different winter wheat backgrounds, Vala, Koútka and Zdar, showed lower winter survival by 20, 36, and 41 % for substitutions of 5B, 5A and 5D, respectively, compared to the original winter cultivars. Reciprocal substitution lines between two winter cultivars Mironovskaya 808 and Bezostaya 1 carrying different recessive alleles, vrn-A1, vrn-B1, vrn-D1, did not exhibit a modified induction of frost tolerance, but the duration of good frost tolerance, as well as the ability to survive the whole winter, was changed. In accordance with the model suggesting that genes for vernalization act as a master switch regulating the duration of frost tolerance, substitutions of homoeologous group 5 chromosomes induced, at first, frost tolerance at a level equal to the parental cultivar, and then, relative to the different extent of saturation of vernalization requirement, they gradually lost both frost tolerance and their ability to re-induce significant frost tolerance with a drop in temperature following warm periods in the winter.     

Effect of the 5R(5A) alien chromosome substitution on the growth habit and winter hardiness of wheat

The growth habit, ear emergence time, and frost tolerance of wheat/rye substitution lines have been studied in cultivars Rang and Mironovskaya Krupnozernaya whose chromosome 5A is substituted with chromosome 5R of Onkhoyskaya rye. Hybrid analysis has demonstrated that the spring habit of the recipient cultivars Rang and Mironovskaya Krupnozernaya is controlled by dominant gene Vrn-A1 located in chromosome 5A. Onokhoyskaya rye has a dominant gene for the spring habit (Sp1) located in chromosome 5R. It has been found that the resultant 5R(5A) alien-substitution lines have a winter type of development and ears do not emerge during summer in plants sown in spring. The change in growth habit has been shown to be related to the absence of the rye Spl gene expression in the substitution lines. The winter hardiness of winter 5R(5A) alien-substitution lines has been studied under the environmental conditions of Novosibirsk. Testing the lines in the first winter demonstrated that their winter survival is 20-27%. The possible presence of the frost resistance gene homeoallelic to the known genes Fr1 and Fr2 of the common wheat located on chromosomes 5A and 5D, respectively, is discussed.     

New data on the distribution of hybrid necrosis genes in winter bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) cultivars

Hybrid necrosis genotypes have been identified in 125 Russian cultivars of winter bread wheat. More than half of them (56%) carry the Ne2 gene (genotype ne1ne1Ne2Ne2); others are free of necrosis genes (genotype ne1ne1ne2ne2). The possible causes of the increase in the Ne2 allele frequency and the loss of the Ne1Ne1ne2ne2 genotype in modern Russian cultivars of winter wheat are discussed. The principal component method has been used to compare the structures of the genetic diversity of cultivars differing in the hybrid necrosis genotype. It has been found that the Ne2 allele in winter wheat cultivars from northern Russia has originated from the cultivar Mironovskaya 808, whereas the cultivar Bezostaya 1 is not a source of this gene. In cultivars from southern Russia, the presence of the Ne2 allele is also mainly accounted for by the use of Mironovskaya 808 wheat in their breeding. The recessive genotype is explained by the presence of descendants of the cultivar Odesskaya 16 in the pedigrees of southern Russian winter wheats. The genetic relationship of cultivars with identical and different necrosis genotypes has been analyzed in nine regions of the Russian Federation.     

Chromosomal location of genes for vernalization requirement duration (<Emphasis Type="Italic">Vrd</Emphasis>) in winter bread wheat

In this study, we carried out monosomic analysis of two isogenic lines of cultivar Mironovskaya 808, monogenically dominant for genes Vrd1 and Vrd2 reducing the cultivar vernalization requirement duration from 50 to respectively 35 and 45 days, as well as analysis of substitution lines Cappelle Desperez/Bezostaya 1. Gene Vrd1 is localized on chromosome 4A; Vrd2, on chromosome 5D. The third gene, which is not allelic to the former two, is present in cultivar Cappelle Desperez on either the 1A, 6A, or 4B chromosome.     

Effect of the 5R(5A) Alien Chromosome Substitution on the Growth Habit and Winter Hardiness of Wheat

The growth habit, ear emergence time, and frost tolerance of wheat/rye substitution lines have been studied in cultivars Rang and Mironovskaya Krupnozernaya whose chromosome 5A is substituted with chromosome 5R of Onkhoyskaya rye. Hybrid analysis has demonstrated that the spring habit of the recipient cultivars Rang and Mironovskaya Krupnozernaya is controlled by dominant gene Vrn-A1 located in chromosome 5A. Onokhoyskaya rye has a dominant gene for the spring habit (Sp1) located in chromosome 5R. It has been found that the resultant 5R(5A) alien-substitution lines have a winter type of development and ears do not emerge during summer in plants sown in spring. The change in growth habit has been shown to be related to the absence of the rye Sp1 gene expression in the substitution lines. The winter hardiness of winter 5R(5A) alien-substitution lines has been studied under the environmental conditions of Novosibirsk. Testing the lines in the first winter demonstrated that their winter survival is 20–27%. The possible presence of the frost resistance gene homeoallelic to the known genes Fr1 and Fr2 of the common wheat located on chromosomes 5A and 5D, respectively, is discussed.     

Relationships among vernalization, shoot apex development and frost tolerance in wheat

BACKGROUND AND AIMS: Frost tolerance of wheat depends primarily upon a strong vernalization requirement, delaying the transition to the reproductive phase. The aim of the present study was to learn how saturation of the vernalization requirement and apical development stage are related to frost tolerance in wheat. METHODS: 'Mironovskaya 808', a winter variety with a long vernalization requirement, and 'Leguan', a spring variety without a vernalization requirement, were acclimated at 2 degrees C at different stages of development. Plant development (morphological stage of the shoot apex), vernalization requirement (days to heading) and frost tolerance (survival of the plants exposed to freezing conditions) were evaluated. KEY RESULTS: 'Mironovskaya 808' increased its frost tolerance more rapidly; it reached a higher level of tolerance and after a longer duration of acclimation at 2 degrees C than was found in 'Leguan'. The frost tolerance of 'Mironovskaya 808' decreased and its ability to re-acclimate a high tolerance was lost after saturation of its vernalization requirement, but before its shoot apex had reached the double-ridge stage. The frost tolerance of 'Leguan' decreased after the plants had reached the floret initiation stage. CONCLUSIONS: The results support the hypothesis that genes for vernalization requirement act as a master switch regulating the duration of low temperature induced frost tolerance. In winter wheat, due to a longer vegetative phase, frost tolerance is maintained for a longer time and at a higher level than in spring wheat. After the saturation of vernalization requirement, winter wheat (as in spring wheat) established only a low level of frost tolerance.     

Location of a gene regulating cold-induced carbohydrate production on chromosome 5A of wheat

 Major changes in osmotic potential during cold acclimation are due to changes in sugar concentration, and there is a good correlation between sugar content and frost tolerance. The objective of the present study was to localize a gene(s) responsible for carbohydrate accumulation during cold acclimation on chromosome 5A of wheat using recombinant lines developed from the cross between the substitution lines Chinese Spring (Cheyenne 5A) and CS(Triticum spelta 5A). Previously, major genes influencing frost resistance (Fr1) and vernalization requirement (Vrn1) had been localized on the long arm of that chromosome. The T. spelta 5A chromosome carrying the Fr1 (frost-sensitive) allele for frost tolerance and the Vrn1 (spring-habit) allele for vernalization requirement did not have a major effect on the sucrose and fructan contents in the Chinese Spring background. On the other hand, the presence of Cheyenne alleles for vernalization requirement, vrn1, and frost tolerance, fr1, significantly increased sugar concentrations. A recombinant line thought to exhibit recombination between the Vrn1 and Fr1 loci suggested that the gene regulating sucrose accumulation was closely associated with, or else represented a pleiotropic effect of, Vrn1, but was separable from the Fr1 locus. Received: 3 March 1997 / Accepted: 7 March 1997     

Differences in the genetic systems controlling photoreaction and vernalization requirements in winter wheat]

Comparing of vernalization requirement and photoperiodic sensitivity in various winter genotypes and F4 lines was carried out. The results of genetic analysis of Ppd genotypes in the crosses of winter wheat cultivars differing in photosensitivity and duration of vernalization requirement are described. It has been shown that differences in duration of vernalization requirement are under control of independent genetic system, distinctive from Ppd gene system regulating level of photosensitivity. This independence does not deny the opposite possibility of Ppd genes to modify duration of vernalization requirement in winter wheat.     

Differences in leaf proteome response to cold acclimation between Lolium perenne plants with distinct levels of frost tolerance

Perennial ryegrass (Lolium perenne) is a high quality forage and turf grass mainly due to its excellent nutritive values and rapid establishment rate. However, this species has limited ability to perform in harsh winter climates. Though winter hardiness is a complex trait, it is commonly agreed that frost tolerance (FT) is its main component. Species growing in temperate regions can acquire FT through exposure to low, non-lethal temperatures, a phenomenon known as cold acclimation (CA). The research on molecular basis of FT has been performed on the model plants, but they are not well adapted to extreme winter climates. Thus, the mechanisms of cell response to low temperature in winter crops and agronomically important perennial grasses have yet to be revealed. Here, two L. perenne plants with contrasting levels of FT, high frost tolerant (HFT) and low frost tolerant (LFT) plants, were selected for comparative proteomic research. The work focused on analyses of leaf protein accumulation before and after 2, 8, 26 h, and 3, 5, 7, 14 and 21 days of CA, using a high-throughput two-dimensional electrophoresis, and on the identification of proteins which were accumulated differentially between the selected plants by the application of mass spectrometry (MS). Analyses of 580 protein profiles revealed a total of 42 (7.2%) spots that showed at a minimum of 1.5-fold differences in protein abundance, at a minimum of at one time point of CA between HFT and LFT genotypes. It was shown that significant differences in profiles of protein accumulation between the analyzed plants appeared most often on the 5th (18 proteins) and the 7th (19 proteins) day of CA. The proteins derived from 35 (83.3%) spots were successfully identified by the use of MS and chloroplast proteins were shown to be the major group selected as differentially accumulated during CA. The functions of the identified proteins and their probable influence on the level of FT in L. perenne are discussed.     

Dynamics of genetic diversity in winter common wheat Triticum aestivum L. cultivars released in Russia from 1929 to 2005

Genealogical analysis was used to study the dynamics of genetic diversity in Russian cultivars of winter common wheat from 1929 to 2005. The Shannon diversity index of the total set of released cultivars remained almost unchanged, although the number of original ancestors (landraces and genetic lines) increased almost tenfold in the period under study. This was explained in terms of the dependence of the modified Shannon diversity index on two parameters, the number of original ancestors and the mean coefficient of parentage. Significant direct effects were revealed: a positive effect of the former parameter and a negative of the latter. As a result, the increase in the number of original ancestors was compensated by the increase in relatedness of cultivars. Genetic erosion of realized diversity was observed, as a half of Russian landraces were lost. Although the mean coefficient of parentage did not reach its critical value (R = 0.25), cultivars of some regions (Central and Volga-Vyatka) proved to be closely related. A favorable gradual decrease in the mean coefficient of parentage was observed in the past 15 years. A set of modem winter wheat cultivars, which were introduced in the Russian State Catalog from 2002 to 2005, displayed a cluster structure. The overwhelming majority of cultivars formed two clusters originating from Bezostaya 1 (67% of cultivars) and Mironovskaya 808 (31%).     

Developmental regulation of metabolites and low temperature tolerance in lines of crosses between spring and winter wheat

Low temperature (LT) tolerance in cereals needs developmental regulation of metabolites, a process which is associated with vernalization requirement. This study was initiated to investigate the relationships among stage of phenological development, final leaf number (FLN), the activities superoxide dismutase, catalase, guaiacol peroxidase, ascorbate peroxidase and polyphenol oxidase, the contents of proline, photosynthetic pigments, and hydrogen peroxide (H₂O₂) during vernalization and LT acclimation in spring and winter wheat. Six genotypes with different vernalization requirements were grown under greenhouse and field conditions. The spring-habit parent, “Pishtaz” and line 4021, rapidly entered the reproductive phase and had a limited capacities to LT acclimate. They also had the lowest antioxidative activities and accumulation of proline among genotypes. Lines 4002 and 4014, with a short vernalization requirement and higher FLN, remained in the early stages of phenological development longer and developed a higher level of LT tolerance and metabolites compared to spring habit genotypes. In contrast, the winter habit “Norstar” and line 4023 spent a longer time in the vegetative stage and accumulated higher levels of metabolites. Maximum LT tolerance and metabolite accumulations occurred near the vegetative/reproductive transition in all genotypes. The longer periods of vernalization and increased FLN that happened along with increased defense mechanisms and decreased damage indices (H₂O₂ content and LT₅₀) ensured LT tolerance in wheat. These results demonstrate that both genetic and environmental factors via developmental regulation of metabolites play important roles in creating LT tolerance in long mild winters of Iran. Significant correlations coefficients for many of the metabolites considered in this study and Lethal temperature 50 (LT₅₀) also suggest that they could be useful as indirect measures of plant LT tolerance potential in wheat breeding programs.     

Dynamics of genetic diversity in winter common wheat Triticum aestivum L. cultivars released in Russia from 1929 to 2005

Genealogical analysis was used to study the dynamics of genetic diversity in Russian cultivars of winter common wheat from 1929 to 2005. The Shannon diversity index of the total set of released cultivars remained almost unchanged, although the number of original ancestors (landraces and genetic lines) increased almost tenfold in the period under study. This was explained in terms of the dependence of the modified Shannon diversity index on two parameters, the number of original ancestors and the mean coefficient of parentage. Significant direct effects were revealed: a positive effect of the former parameter and a negative of the latter. As a result, the increase in the number of original ancestors was compensated by the increase in relatedness of cultivars. Genetic erosion of released diversity was observed, as a half of Russian landraces were lost. Although the mean coefficient of parentage did not reach its critical value $(\bar R = 0.25)$ , cultivars of some regions (Central and Volga-Vyatka) proved to be closely related. A favorable gradual decrease in the mean coefficient of parentage was observed in the past 15 years. A set of modern winter wheat cultivars, which were introduced in the Russian Official List from 2002 to 2005, displayed a cluster structure. The overwhelming majority of cultivars formed two clusters originating from Bezostaya 1 (67% of cultivars) and Mironovskaya 808 (31%).