Epistatic interaction between vernalization genes Vrn-Am1 and Vrn-Am2 in diploid wheat

Genes Vrn-A(m)1 and Vrn-A(m)2 control the vernalization requirement in diploid wheat (Triticum monococcum). The epistatic interaction between these two genes on flowering date was studied here using a factorial analysis of variance. One hundred and two F2 plants were classified according to their genotypes for molecular markers tightly linked to Vrn-A(m)1 and Vrn-A(m)2. Mean comparisons showed that the VrnA(m)2 allele for winter growth habit was dominant to the vrn-A(m)2 allele for spring growth habit and that the Vrn-A(m)1 allele for spring growth habit was dominant to the vrn-A(m)1 allele for winter growth habit. A significant interaction was found between these two genes, suggesting that they work in the same developmental pathway. Plants homozygous for the recessive vrn-A(m)2 allele for spring growth habit flowered earlier than plants from the Vrn-A(m)2 class independently of the alleles present at Vrn-A(m)1. However, differences in heading date between plants with the Vrn-A(m)1 allele and those with the vrn-A(m)1 allele were significant only when the dominant Vrn-A(m)2 allele was present. A genetic model for the action of these two vernalization genes is proposed in which the role of Vrn-A(m)1 is to counteract the Vrn-A(m)2-mediated delay of flowering.     

Developmental traits affecting low-temperature tolerance response in near-isogenic lines for the Vernalization locus Vrn-A1 in wheat (Triticum aestivum L. em Thell)

Investigation of low-temperature (LT) tolerance in cereals has commonly led to the region of the vyn-A1 vernalization gene or its homologue in related genomes. Two cultivars, one a non-hardy spring wheat and one a very cold-hardy winter wheat, whose growth habits are determined by the Vrn-A1 (spring habit) and vrn-A1 (winter habit) alleles, were chosen to produce reciprocal near-isogenic lines (NILs). These lines were then used to determine the relationship between rate of phenological development and the degree and duration of LT tolerance gene expression. Each allele was isolated in the genetic backgrounds of the non-hardy spring wheat 'Manitou' and the very cold-hardy winter wheat 'Norstar'. The effects of each allele on phenological development and low-temperature tolerance (LT50) were determined at regular intervals over a 4 degrees C acclimation period of 0-98 d. The vegetative/reproductive transition, as determined by final leaf number (FLN), was found to be a major developmental factor influencing LT tolerance. Possession of a vernalization requirement increased both the length of the vegetative growth phase and LT tolerance. Similarly, increased FLN in spring Norstar and winter Manitou NILs delayed their vegetative/reproductive transition and increased their LT tolerance relative to Manitou. Although the winter Manitou NILs had a lower FLN than the spring Norstar NILs, they were able to extend their vegetative stage to a similar length by increasing the phyllochron (interval between the appearance of successive leaves). Cereal plants have four ways of increasing the length of the vegetative phase, all of which extend the time that low-temperature tolerance genes are more highly expressed: (1) vernalization; (2) photoperiod responses; (3) increased leaf number; and (4) increased length of the phyllochron.     

Identification of quantitative trait loci and associated candidate genes for low-temperature tolerance in cold-hardy winter wheat

Low-temperature (LT) tolerance is an important economic trait in winter wheat (Triticum aestivum L.) that determines the plants’ ability to cope with below freezing temperatures. Essential elements of the LT tolerance mechanism are associated with the winter growth habit controlled by the vernalization loci (Vrn-1) on the group 5 chromosomes. To identify genomic regions, which in addition to vrn-1 determine the level of LT tolerance in hexaploid wheat, two doubled haploid (DH) mapping populations were produced using parents with winter growth habit (vrn-A1, vrn-B1, and vrn-D1) but showing different LT tolerance levels. A total of 107 DH lines were analyzed by genetic mapping to produce a consensus map of 2,873 cM. The LT tolerance levels for the Norstar (LT₅₀=−20.7°C) × Winter Manitou (LT₅₀=−14.3°C) mapping population ranged from −12.0 to −22.0°C. Single marker analysis and interval mapping of phenotyped lines revealed a major quantitative trait locus (QTL) on chromosome 5A and a weaker QTL on chromosome 1D. The 5A QTL located 46 cM proximal to the vrn-A1 locus explained 40% of the LT tolerance variance. Two C-repeat Binding Factor (CBF) genes expressed during cold acclimation in Norstar were located at the peak of the 5A QTL.     

The Characterization and Geographical Distribution of the Genes Responsible for Vernalization Requirement in Chinese Bread Wheat

The frequency and distribution of the major vernalization requirement genes and their effects on growth habits were studied.Of the 551 bread wheat genotypes tested,seven allelic combinations of the three Vrn.1 genes were found to be responsible for the spring habit,three for the facultative habit and one for the winter habit.The three Vrn-1 genes behaved additively with the dominant allele of Vrn-A1 exerting the strongest effect.The allele combinations of the facultative genotypes and the discovery of spring genotypes with "winter" allele of Vrn-1 implied the presence of as yet unidentified alleles/genes for vernalization response.The dominant alleles of the three Vrn-1 genes were found in all ten ecological regions where wheat Is cultivated in China,with Vrn-D1 as the most common allele in nine and Vrn-A1 in one.The combination of vrn-A 1vrnB 1Vrn-D1 was the predominant genotype in seven of the regions.Compared with landraces,improved varieties contain a higher proportion of the spring type.This was attributed by a higher frequency of the dominant Vrn-A1 and Vrn-B1 alleles in the latter.Correlations between Vrn-1 allelic constitutions and heading date,spike length,plant type as well as cold tolerance were established.     

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.     

Validation of the <Emphasis Type="Italic">VRN-H2</Emphasis>/<Emphasis Type="Italic">VRN-H1</Emphasis> epistatic model in barley reveals that intron length variation in <Emphasis Type="Italic">VRN-H1</Emphasis> may account for a continuum of vernalization sensitivity

The epistatic interaction of alleles at the VRN-H1 and VRN-H2 loci determines vernalization sensitivity in barley. To validate the current molecular model for the two-locus epistasis, we crossed homozygous vernalization-insensitive plants harboring a predicted “winter type” allele at either VRN-H1 (Dicktoo) or VRN-H2 (Oregon Wolfe Barley Dominant), or at both VRN-H (Calicuchima-sib) loci and measured the flowering time of unvernalized F₂ progeny under long-day photoperiod. We assessed whether the spring growth habit of Calicuchima-sib is an exception to the two-locus epistatic model or contains novel “spring” alleles at VRN-H1 (HvBM5A) and/or VRN-H2 (ZCCT-H) by determining allele sequence variants at these loci and their effects relative to growth habit. We found that (a) progeny with predicted “winter type” alleles at both VRN-H1 and VRN-H2 alleles exhibited an extremely delayed flowering (i.e. vernalization-sensitive) phenotype in two out of the three F₂ populations, (b) sequence flanking the vernalization critical region of HvBM5A intron 1 likely influences degree of vernalization sensitivity, (c) a winter habit is retained when ZCCT-Ha has been deleted, and (d) the ZCCT-H genes have higher levels of allelic polymorphism than other winterhardiness regulatory genes. Our results validate the model explaining the epistatic interaction of VRN-H2 and VRN-H1 under long-day conditions, demonstrate recovery of vernalization-sensitive progeny from crosses of vernalization-insensitive genotypes, show that intron length variation in VRN-H1 may account for a continuum of vernalization sensitivity, and provide molecular markers that are accurate predictors of “winter vs spring type” alleles at the VRN-H loci.     

Large deletions within the first intron in <Emphasis Type="Italic">VRN-1</Emphasis> are associated with spring growth habit in barley and wheat

The broad adaptability of wheat and barley is in part attributable to their flexible growth habit, in that spring forms have recurrently evolved from the ancestral winter growth habit. In diploid wheat and barley growth habit is determined by allelic variation at the VRN-1 and/or VRN-2 loci, whereas in the polyploid wheat species it is determined primarily by allelic variation at VRN-1. Dominant Vrn-A1 alleles for spring growth habit are frequently associated with mutations in the promoter region in diploid wheat and in the A genome of common wheat. However, several dominant Vrn-A1, Vrn-B1, Vrn-D1 (common wheat) and Vrn-H1 (barley) alleles show no polymorphisms in the promoter region relative to their respective recessive alleles. In this study, we sequenced the complete VRN-1 gene from these accessions and found that all of them have large deletions within the first intron, which overlap in a 4-kb region. Furthermore, a 2.8-kb segment within the 4-kb region showed high sequence conservation among the different recessive alleles. PCR markers for these deletions showed that similar deletions were present in all the accessions with known Vrn-B1 and Vrn-D1 alleles, and in 51 hexaploid spring wheat accessions previously shown to have no polymorphisms in the VRN-A1 promoter region. Twenty-four tetraploid wheat accessions had a similar deletion in VRN-A1 intron 1. We hypothesize that the 2.8-kb conserved region includes regulatory elements important for the vernalization requirement. Epistatic interactions between VRN-H2 and the VRN-H1 allele with the intron 1 deletion suggest that the deleted region may include a recognition site for the flowering repression mediated by the product of the VRN-H2 gene of barley.     

Molecular and Structural Characterization of Barley Vernalization Genes

Vernalization, the requirement of a period of low temperature to induce transition from the vegetative to reproductive state, is an evolutionarily and economically important trait in the Triticeae. The genetic basis of vernalization in cultivated barley (Hordeum vulgare subsp. vulgare) can be defined using the two-locus VRN-H1/VRN-H2 model. We analyzed the allelic characteristics of HvBM5A, the candidate gene for VRN-H1, from ten cultivated barley accessions and one wild progenitor accession (subsp. spontaneum), representing the three barley growth habits – winter, facultative, and spring. We present multiple lines of evidence, including sequence, linkage map location, and expression, that support HvBM5A being VRN-H1. While the predicted polypeptides from different growth habits are identical, spring accessions contain a deletion in the first intron of HvBM5A that may be important for regulation. While spring HvBM5A alleles are typified by the intron-localized deletion, in some cases, the promoter may also determine the allele type. The presence/absence of the tightly linked ZCCT-H gene family members on chromosome 4H perfectly correlates with growth habit and we conclude that one of the three ZCCT-H genes is VRN-H2. The VRN-H2 locus is present in winter genotypes and deleted from the facultative and spring genotypes analyzed in this study, suggesting the facultative growth habit (cold tolerant, vernalization unresponsive) is a result of deletion of the VRN-H2 locus and presence of a winter HvBM5A allele. All reported barley vernalization QTLs can be explained by the two-locus VRN-H1/VRN-H2 model based on the presence/absence of VRN-H2 and a winter vs. spring HvBM5A allele. Electronic Supplementary Material Electronic Supplementary material is available for this article at and accessible for authorised users.     

Allelic variation at the linked AP1 and PhyC loci in hexaploid wheat is associated but not perfectly correlated with vernalization response

Vernalization requirement is an important trait in temperate crop plants such as wheat and must be considered when selecting varieties for cultivation under different climatic conditions. To determine the growth habit of wheat varieties, plants need to be grown under different vernalization regimes, a lengthy but necessary process for breeders involved in crossing winter with spring germplasm. If haplotypes can be associated with growth habit, then molecular marker assays that are reliable, cheap, and quick can be developed to assist in the selection of plants with the desired phenotype. We have analyzed 81 accessions that have different vernalization requirements and putative different origins of spring habit for sequence variation at the Apetala1 (AP1) locus, which underlies Vrn-1, and at the linked Phytochrome C (PhyC) locus. Good correspondence was found between the AP1 genotype and the PhyC haplotype for 77 of the 81 accessions. Two varieties displayed a recombination event between the AP1 and PhyC loci, and one variety carried a recombinant PhyC gene. In addition, one variety carried an apparent AP1 winter allele, but displayed the Vrn-A1 spring habit. The PhyC haplotype for this variety also indicated the presence of a Vrn-A1 spring allele. Our data suggest that both the AP1 promoter region and PhyC SNPs can be used as diagnostic markers for vernalization response at the vrn-A1 locus, but that neither are perfect tags.     

Genetic loci associated with stem elongation and winter dormancy release in wheat

In winter wheat (Triticum aestivum L.), the stem begins to elongate after the vernalization requirement is satisfied during winter and when favorable temperature and photoperiod conditions are attained in spring. In this study, we precisely measured elongation of the first extended internode on 96 recombinant inbred lines of a population that was generated from a cross between two winter wheat cultivars, Jagger (early stem elongation) and 2174 (late stem elongation). We mapped a major locus for stem elongation to the region where VRN-A1 resides in chromosome 5A. Visible assessment of winter dormancy release was concomitantly associated with this locus. VRN1 was previously cloned based on variation in vernalization requirement between spring wheat carrying a dominant Vrn-1 allele and winter wheat carrying a recessive vrn-1 allele. Both of two winter wheat cultivars in this study carry a recessive vrn-A1 allele; therefore, our results suggest that either VRN-A1 might invoke a new regulatory mechanism or a new gene residing close to VRN-A1 plays a regulatory role in winter wheat development. Phenotypic expression of the vrn-A1a allele of Jagger was more sensitive to the year of measurement of stem elongation than that of the vrn-A1b allele of 2174. In addition to QSte.osu.5A, several loci were also found to have minor effects on initial stem elongation of winter wheat. Seventeen of nineteen locally adapted cultivars in the southern Great Plaints contained the vrn-A1b allele. Hence, breeders in this area have inadvertently selected this allele, contributing to later stem elongation and more conducive developmental patterns for grain production.     

The relationship between vernalization-and photoperiodically-regulated genes and the development of frost tolerance in wheat and barley

The review summarizes the level of current knowledge of impacts of vernalization and photoperiod on the induction and maintenance of frost tolerance (FrT) in wheat and barley. The phenomenon of vernalization is briefly described and the major vernalization (VRN) loci are characterised. Vernalization requirement and the three major growth habits of Triticeae (facultative, winter and spring) are defined on the basis of the two-locus VRN-2/VRN-1 epistatic model. Major photoperiodically regulated genes, which influence the transition to flowering, are characterised and their interactions with VRN genes are briefly discussed. The phenomenon of induction of FrT during the process of cold acclimation (CA) is described and the major cold-induced Cor/Lea genes are listed. Important regulatory mechanisms, i.e., CBF pathway, controlling the expression of Cor/Lea genes under cold, are discussed. The major loci affecting the development of FrT in Triticeae, the Fr loci, are characterised. In conclusion, current progress in this research field is summarized and new questions arising in the area are formulated.     

Genes responding to vernalization in hexaploid wheat

Genotype-specific gene expression in response to vernalization in common wheat was examined by the differential display method. Two near-isogenic lines of Vrn-A1 ( Vrn-A1 for the spring type and vrn-A1 for the winter type) were treated by vernalization of developing embryos in detached-ear cultures. This treatment was effective to promote vrn-A1 genotypes to head at a time equivalent to that of Vrn-A1. Differential cDNA fragments were isolated by the RT-PCR method from embryos subjected to vernalization treatments for 2- and 4-weeks at DAP10 and DAP20 stages, respectively. Among 110 differential cDNA fragments isolated, 48 were examined for their chromosomal locations and designated as wec ( wheat- embryo cold treatment) genes. Seven wec genes showed genotype-specific expression in response to vernalization. The statistical analysis utilizing two recombinant inbred lines showed that four wec genes were significantly associated with heading factors.     

Low-temperature acclimation of barley cultivars used as parents in mapping populations: response to photoperiod,vernalization and phenological development

Six barley (Hordeum vulgare L.) accessions, previously used as parents of mapping populations, were evaluated for characters potentially affecting the location of low-temperature (LT) tolerance QTLs. Three were of winter growth habit (Kompolti Korai, Nure, and Strider), one was facultative (Dicktoo) and two were spring (Morex and Tremois). Final leaf number (FLN) and LT₅₀ were determined at weekly intervals from 0 to 98 days of LT acclimation/vernalization under both long day (LD) and short day (SD) photoperiods. The point of vegetative/reproductive transition was determined from measurements of double ridge (DR) formation and FLN. With the exception of Nure, SD delayed development by increasing leaf production. Dicktoo was extremely SD sensitive lengthening its vegetative phase by more than 63 days relative to the LD photoperiod. SD had the opposite effect on Nure, causing an accelerating of flowering exhibiting the characteristic of ‘short day vernalization’. All accessions except Dicktoo and Kompolti Korai acclimated rapidly in the first 7 days of LT exposure, approaching their maximum LT tolerance in 14–21 days. Dicktoo and Kompolti Korai continued to slowly acclimate until reproductive transition. The results emphasize two important points: (1) the location of QTLs for LT tolerance, and as a consequence the identification of putative candidate genes, will be a function of the genotypes sampled, the experimental conditions used, and the quality of the phenotypic data and (2) the barley LT tolerance pathway reaches an early impediment relative to closely related more hardy members of the Triticeae such as wheat and rye.     

Genetics of flowering time in bread wheat <Emphasis Type="Italic">Triticum aestivum</Emphasis>: complementary interaction between vernalization-insensitive and photoperiod-insensitive mutations imparts very early flowering habit to spring wheat

Time to flowering in the winter growth habit bread wheat is dependent on vernalization (exposure to cold conditions) and exposure to long days (photoperiod). Dominant Vrn-1 (Vrn-A1, Vrn-B1 and Vrn-D1) alleles are associated with vernalization independent spring growth habit. The semidominant Ppd-D1a mutation confers photoperiod-insensitivity or rapid flowering in wheat under short day and long day conditions. The objective of this study was to reveal the nature of interaction between Vrn-1 and Ppd-D1a mutations (active alleles of the respective genes vrn-1 and Ppd-D1b). Twelve Indian spring wheat cultivars and the spring wheat landrace Chinese Spring were characterized for their flowering times by seeding them every month for five years under natural field conditions in New Delhi. Near isogenic Vrn-1 Ppd-D1 and Vrn-1 Ppd-D1a lines constructed in two genetic backgrounds were also phenotyped for flowering time by seeding in two different seasons. The wheat lines of Vrn-A1a Vrn-B1 Vrn-D1 Ppd-D1a, Vrn-A1a Vrn-B1 Ppd-D1a and Vrn-A1a Vrn-D1 Ppd-D1a (or Vrn-1 Ppd-D1a) genotypes flowered several weeks earlier than that of Vrn-A1a Vrn-B1 Vrn-D1 Ppd-D1b, Vrn-A1b Ppd-D1b and Vrn-D1 Ppd-D1b (or Vrn-1 Ppd-D1b) genotypes. The flowering time phenotypes of the isogenic vernalization-insensitive lines confirmed that Ppd-D1a hastened flowering by several weeks. It was concluded that complementary interaction between Vrn-1 and Ppd-D1a active alleles imparted super/very-early flowering habit to spring wheats. The early and late flowering wheat varieties showed differences in flowering time between short day and long day conditions. The flowering time in Vrn-1 Ppd-D1a genotypes was hastened by higher temperatures under long day conditions. The ambient air temperature and photoperiod parameters for flowering in spring wheat were estimated at 25°C and 12 h, respectively.     

Allelic variation at the <Emphasis Type="Italic">VRN-1</Emphasis> promoter region in polyploid wheat

Vernalization, the requirement of a long exposure to low temperatures to induce flowering, is an essential adaptation of plants to cold winters. We have shown recently that the vernalization gene VRN-1 from diploid wheat Triticum monococcum is the meristem identity gene APETALA1, and that deletions in its promoter were associated with spring growth habit. In this study, we characterized the allelic variation at the VRN-1 promoter region in polyploid wheat. The Vrn-A1a allele has a duplication including the promoter region. Each copy has similar foldback elements inserted at the same location and is flanked by identical host direct duplications (HDD). This allele was found in more than half of the hexaploid varieties but not among the tetraploid lines analyzed here. The Vrn-A1b allele has two mutations in the HDD region and a 20-bp deletion in the 5 UTR compared with the winter allele. The Vrn-A1b allele was found in both tetraploid and hexaploid accessions but at a relatively low frequency. Among the tetraploid wheat accessions, we found two additional alleles with 32 bp and 54 bp deletions that included the HDD region. We found no size polymorphisms in the promoter region among the winter wheat varieties. The dominant Vrn-A1 allele from two spring varieties from Afghanistan and Egypt (Vrn-A1c allele) and all the dominant Vrn-B1 and Vrn-D1 alleles included in this study showed no differences from their respective recessive alleles in promoter sequences. Based on these results, we concluded that the VRN-1 genes should have additional regulatory sites outside the promoter region studied here.     

Molecular characterization of vernalization response genes in Canadian spring wheat.

Vernalization response (Vrn) genes play a major role in determining the flowering/maturity times of spring-sown wheat. We characterized a representative set of 40 western Canadian adapted spring wheat cultivars/lines for 3 Vrn loci. The 40 genotypes were screened, along with 4 genotypes of known Vrn genes, using previously published genome-specific polymerase chain reaction primers designed for detecting the presence or absence of dominant or recessive alleles of the major Vrn loci: Vrn-A1, Vrn-B1, and Vrn-D1. The dominant promoter duplication allele Vrn-A1a was present in 34 of 40 cultivars/lines, whereas the promoter deletion allele Vrn-A1b was present in only 1 of the western Canadian cultivars (Triticum aestivum L. 'Rescue') and 2 of its derivative chromosomal substitution lines. The intron deletion allele Vrn-A1c was not present in any line tested. Only 4 of the western Canadian spring wheat cultivars tested here carry the recessive vrn-A1 allele. The dominant allele of Vrn-B1 was detected in 20 cultivars/lines. Fourteen cultivars/lines had dominant alleles of Vrn-A1a and Vrn-B1 in combination. All cultivars/lines carried the recessive allele for Vrn-D1. The predominance of the dominant allele Vrn-A1a in Canadian spring wheat appears to be due to the allele's vernalization insensitivity, which confers earliness under nonvernalizing growing conditions. Wheat breeders in western Canada have incorporated the Vrn-A1a allele into spring wheats mainly by selecting for early genotypes for a short growing season, thereby avoiding early and late season frosts. For the development of early maturing cultivars with high yield potential, different combinations of Vrn alleles may be incorporated into spring wheat breeding programs in western Canada.     

Inheritance of cell size in wheat (Triticum aestivum L.) and its relationship to the vernalization loci

Reduced cell size is an important adaptive feature in plant response to environmental stresses. The objectives of the present study were to determine the inheritance and location of genes controlling cell size and to establish the relationship between cell size, low-temperature (LT) tolerance, and growth habit as determined by the Vrn loci in wheat. Guard cell length was measured in F₁, F₂, andF₂-derived F₃ populations from parents ranging widely in cell size and in the Chinese Spring/ Cheyenne (CS/CNN) chromosome substitution series. The cell size of F₁ hybrids was similar to the parental midpoint and the F₂ frequency distribution was symmetrical about the mean indicating that cell size was determined by additive gene action with little or no dominance. It appears that there are several genes involved since none of the F₂ progeny had a cell size as large or as small as the parental mean range. The cell size of the homozygous spring and winter lines from F₂-derived F₃ populations fell into two distinct groups that were related to plant growth habit. Large cell size was associated with the spring-habit alleles (Vrn-A1) and small cell size was associated with the winter-habit alleles (vrn-A1) on chromosome 5A. Analyses of the CS/CNN chromosome substitution series showed that CNN chromosomes 5A and 5B both reduced cell size without changing the growth habit, indicating that growth habit per se does not determine cell size. The group-5 chromosomes therefore appear to carry homoeologous alleles with major effects on cell size in wheat. This places cell-size control and many other low-temperature (LT) tolerance associated characters in close proximity to the vrn region of the group-5 chromosomes. Received: 17 August 2000 / Accepted: 20 November 2000     

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.     

VRN1 genes variability in tetraploid wheat species with a spring growth habit

Background

Vernalization genes VRN1 play a major role in the transition from vegetative to reproductive growth in wheat. In di-, tetra- and hexaploid wheats the presence of a dominant allele of at least one VRN1 gene homologue (Vrn-A1, Vrn-B1, Vrn-G1 or Vrn-D1) determines the spring growth habit. Allelic variation between the Vrn-1 and vrn-1 alleles relies on mutations in the promoter region or the first intron. The origin and variability of the dominant VRN1 alleles, determining the spring growth habit in tetraploid wheat species have been poorly studied.

Results

Here we analyzed the growth habit of 228 tetraploid wheat species accessions and 25 % of them were spring type. We analyzed the promoter and first intron regions of VRN1 genes in 57 spring accessions of tetraploid wheats. The spring growth habit of most studied spring accessions was determined by previously identified dominant alleles of VRN1 genes. Genetic experiments proof the dominant inheritance of Vrn-A1d allele which was widely distributed across the accessions of Triticum dicoccoides. Two novel alleles were discovered and designated as Vrn-A1b.7 and Vrn-B1dic. Vrn-A1b.7 had deletions of 20 bp located 137 bp upstream of the start codon and mutations within the VRN-box when compared to the recessive allele of vrn-A1. So far the Vrn-A1d allele was identified only in spring accessions of the T. dicoccoides and T. turgidum species. Vrn-B1dic was identified in T. dicoccoides IG46225 and had 11 % sequence dissimilarity in comparison to the promoter of vrn-B1. The presence of Vrn-A1b.7 and Vrn-B1dic alleles is a predicted cause of the spring growth habit of studied accessions of tetraploid species. Three spring accessions T. aethiopicum K-19059, T. turanicum K-31693 and T. turgidum cv. Blancal possess recessive alleles of both VRN-A1 and VRN-B1 genes. Further investigations are required to determine the source of spring growth habit of these accessions.

Conclusions

New allelic variants of the VRN-A1 and VRN-B1 genes were identified in spring accessions of tetraploid wheats. The origin and evolution of VRN-A1 alleles in di- and tetraploid wheat species was discussed.