Winter wheats require several weeks at low temperature to flower. This process, vernalization, is controlled mainly by the VRN1 gene. Using 6,190 gametes, we found VRN1 to be completely linked to MADS-box genes AP1 and AGLG1 in a 0.03-centimorgan interval flanked by genes Cysteine and Cytochrome B5. No additional genes were found between the last two genes in the 324-kb Triticum monococcum sequence or in the colinear regions in rice and sorghum. Wheat AP1 and AGLG1 genes were similar to Arabidopsis meristem identity genes AP1 and AGL2, respectively. AP1 transcription was regulated by vernalization in both apices and leaves, and the progressive increase of AP1 transcription was consistent with the progressive effect of vernalization on flowering time. Vernalization was required for AP1 transcription in apices and leaves in winter wheat but not in spring wheat. AGLG1 transcripts were detected during spike differentiation but not in vernalized apices or leaves, suggesting that AP1 acts upstream of AGLG1. No differences were detected between genotypes with different VRN1 alleles in the AP1 and AGLG1 coding regions, but three independent deletions were found in the promoter region of AP1. These results suggest that AP1 is a better candidate for VRN1 than AGLG1. The epistatic interactions between vernalization genes VRN1 and VRN2 suggested a model in which VRN2 would repress directly or indirectly the expression of AP1. A mutation in the promoter region of AP1 would result in the lack of recognition of the repressor and in a dominant spring growth habit.

Winter wheat and barley varieties require an extended exposure to low temperatures to accelerate flowering (vernalization), whereas spring varieties do not have this requirement. In this study, we show that in these species, the vernalization gene VRN3 is linked completely to a gene similar to Arabidopsis FLOWERING LOCUS T (FT).

Plants with a winter growth habit flower earlier when exposed for several weeks to cold temperatures, a process called vernalization. We report here the positional cloning of the wheat vernalization gene VRN2, a dominant repressor of flowering that is downregulated by vernalization. Loss of function of VRN2, whether by natural mutations or deletions, resulted in spring lines, which do not require vernalization to flower. Reduction of the RNA level of VRN2 by RNA interference accelerated flowering time of transgenic winter wheat plants more than a month.Common wheat (Triticum aestivum L.) is one of the primary grains consumed by humans and is grown in very different environments. This wide adaptability has been favored by the existence of wheat varieties with different growth habits. Winter wheats require a long exposure to low temperatures to flower (vernalization) and are sown in the fall, whereas spring wheats do not have a vernalization requirement and can be planted in spring or fall. The genes from the vernalization pathway prevent flower development during the winter, providing protection for the temperature-sensitive floral organs against the cold.VRN1 and VRN2 are the central genes in the vernalization pathway in wheat, barley, and other temperate cereals. These two genes have strong epistatic interactions and are likely part of the same regulatory pathway (1, 2). In both diploid wheat (Triticum monococcum L.)

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.

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 VrnA1a 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.

No abstract

Wheat is usually classified as a long day (LD) plant because most varieties flower earlier when exposed to longer days. In addition to LD, winter wheats require a long exposure to low temperatures (vernalization) to become competent for flowering. Here we show that in some genotypes this vernalization requirement can be replaced by interrupting the LD treatment by six weeks of short day (SD), and that this replacement is associated with the SD down-regulation of the VRN2 flowering repressor. In addition, we found that SD down-regulation of VRN2 at room temperature is not followed by the up-regulation of the meristem identity gene VRN1 until plants are transferred to LD. This result contrasts with the VRN1 up-regulation observed after the VRN2 down-regulation by vernalization, suggesting the existence of a second VRN1 repressor. Analysis of natural VRN1 mutants indicated that a CArG-box located in the VRN1 promoter is the most likely regulatory site for the interaction with this second repressor. Up-regulation of VRN1 under SD in accessions carrying mutations in the CArG-box resulted in an earlier initiation of spike development, compared to other genotypes. However, even the genotypes with CArG box mutations required LD for a normal and timely spike development. The SD acceleration of flowering was observed in photoperiod sensitive winter varieties. Since vernalization requirement and photoperiod sensitivity are ancestral traits in Triticeae species we suggest that wheat was initially a SD-LD plant and that strong selection pressures during domestication and breeding resulted in the modification of this dual regulation. The down-regulation of the VRN2 repressor by SD is likely part of the mechanism associated with the SD-LD regulation of flowering in photoperiod sensitive winter wheat.

Vernalization, the requirement of a long exposure to low temperatures to accelerate flowering, is an essential adaptation of plants to cold winters. The vernalization gene VRN-1 plays an important role in this process in diploid (Triticum monococcum) and polyploid wheat (Triticum aestivum). We have recently shown that the diploid wheat VRN-A m 1 gene was similar to the Arabidopsis (Arabidopsis thaliana L. Heynh.) APETALA1 meristem identity gene. We also showed that dominant Vrn-A m 1 alleles were the result of loss-of-function mutations in regulatory regions recognized by a VRN-1 repressor, likely VRN-2. This model predicts that only the dominant Vrn-1 allele will be transcribed in lines carrying both recessive and dominant alleles. Here, we confirm this prediction in young isogenic lines of hexaploid wheat carrying different dominant Vrn-A1, Vrn-B1, and Vrn-D1 alleles, and also in heterozygous VRN-1 diploid wheat plants. However, a few weeks later, transcripts from the recessive alleles were also detected in both the polyploid and heterozygous diploid spring plants. Transcription of the recessive alleles was preceded by a reduction of the transcript levels of VRN-2. These results suggest that the dominant Vrn-1 allele or a gene regulated by VRN-1 down-regulates the VRN-2 repressor facilitating the transcription of the recessive alleles in unvernalized plants. We also show here that the level of VRN-1 transcripts in early developmental stages is critical for flowering initiation. A reduction of VRN-1 transcript levels by RNA interference delayed apex transition to the reproductive stage, increased the number of leaves, and delayed heading time by 2 to 3 weeks. We hypothesize that the coordinated transcription of dominant and recessive alleles may contribute to an earlier attainment of the VRN-1 transcript level threshold required to trigger flowering initiation in polyploid wheat.Common wheat (Triticum aestivum) was one of the first crops domesticated by man and, since then, has traveled with human populations to a wide range of environments. Wheat adaptability to this wide range of environments is partially due to the exploitation of genetic variation in daylength sensitivity and vernalization requirement, which provide a flexible regulation of flowering time. Vernalization, the requirement of a prolonged exposure to low temperatures to accelerate flowering, is particularly important for fall-planted wheat varieties to prevent flower development during winter and protect the environmentally sensitive floral organs. Wheat varieties that require vernalization to flower are referred to as winter wheats, whereas those without a vernalization requirement are identified as spring wheats.Allelic variation at the VRN-1 locus is one of the main sources of genetic differences in vernalization requirement in both diploid (Triticum monococcum) and polyploid wheats (for review, see McIntosh et al., 2003). Common wheat is a hexaploid species (2n 5 42, genomes AABBDD) and carries three homoeologous copies of the VRN-1 gene, o...