Vrn-A1 Alleles Research Articles

Allelic Variation of Rht-1, Vrn-1 and Ppd-1 in Korean Wheats and Its Effect on Agronomic Traits

The allelic variations at the Rht-1, Vrn-1 and Ppd-1 of 410 Korean wheat cultivars, including 111 Korean experimental lines, 238 Korean landraces and 61 North Korean collections, were investigated to provide the information of plant height and heading date and to elucidate the relationship between those traits and allelic variation of these genes because earliness is major consideration in Korean wheat production. All Korean wheats displayed vrn-A1 and Ppd-A1b alleles, while Rht-B1a, Rht-D1a, vrn-B1, Vrn-D1, Ppd-B1b and Ppd-D1a alleles were also predominantly found. Most Korean wheats carried both Rht-B1a and Rht-D1a alleles, both vrn-B1 and Vrn-D1 alleles, or both Ppd-B1b and Ppd-D1a alleles. The Rht-B1a, vrn-D1, Ppd-B1b and Ppd-D1b alleles were found to exhibit longer culm and spike length than their counterpart alleles. The Rht-B1a allele also showed longer spike length than Rht-B1b. Vrn-B1b and vrn-D1 alleles exhibited longer days to heading date than their counterpart alleles at the Vrn-B1 and Vrn-D1 loci. Lines carrying both Rht-B1b and Rht-D1b alleles displayed shorter culm and longer spike length and days to heading date than any other combination of alleles at the Rht-B1 and Rht-D1 loci. In contrast, lines carrying both Ppd-B1b and Ppd-D1b alleles exhibited longer culm and spike length than any other combination of alleles at the Ppd-B1 and Ppd-D1 loci.

Study of allelic composition of Vrn-1 and Ppd-1 genes in early–ripening and middle–early varieties of spring soft wheat in siberia

Using allele-specific primers, a combination of the Vrn-A1, Vrn-B1, Vrn-D1, Ppd-D1, and Ppd-B1 alleles was analyzed in 48 early-ripening and middle–early varieties of the spring soft wheat from different breeding centers of Siberia. Six haplotypes were detected, including two most widespread with two dominant genes (Vrn-A1 and Vrn-B1) on the background of recessivePpd-D1b gene (causing sensitivity to the photoperiod). The dominant Ppd-D1a allele was found in combination with dominant Vrn-A1 and Vrn-B1 genes only in one variety (Tulun 15), which is the earliest among all the analyzed varieties. A significant variability in the duration of vegetation was detected within each haplotype assuming a strong influence of the genetic background on the given trait. The results obtained can be used in marker-assisted selection of the most optimal genotypes for specific cultivation conditions.

VRN-1 gene- associated prerequisites of spring growth habit in wild tetraploid wheat T. dicoccoides and the diploid A genome species.

BackgroundIn order to clarify the origin of spring growth habit in modern domesticated wheat, allelic variability of the VRN-1 gene was investigated in a wide set of accessions of the wild tetraploid species Triticum dicoccoides (BBAA), together with diploid species T. monococcum, T. boeoticum and T. urartu, presumable donors of the A genome to polyploid wheats.ResultsNo significant variation was found at the VRN-B1 locus of T. dicoccoides, whereas at VRN-A1 a number of previously described alleles were found with small deletions in the promoter (VRN-A1b, VRN-A1d) or a large deletion in the first (1st) intron (VRN-A1L). The diploid A genome species were characterized by their own set of VRN-1 alleles including previously described VRN-A1f and VRN-A1h alleles with deletions in the promoter region and the VRN-A1ins allele containing a 0.5 kb insertion in the 1st intron. Based on the CAPS screening data, alleles VRN-A1f and VRN-A1ins were species-specific for T. monococcum, while allele VRN-A1h was specific for T. boeoticum. Different indels were revealed in both the promoter and 1st intron of the recessive VRN-A1u allele providing specific identification of T. urartu, the proposed donor of the A genome to modern wheat. We found that alleles VRN-A1b and VRN-A1h, previously described as dominant, have either no or weak association with spring growth habit, while in some diploid accessions this habit was associated with the recessive VRN-A1 allele.ConclusionsSpring growth habit in diploid wheats was only partially associated with indels in regulatory regions of the VRN-1 gene. An exception is T. monococcum where dominant mutations in both the promoter region and, especially, the 1st intron were selected during domestication resulting in a greater variety of spring forms. The wild tetraploid T. dicoccoides had a distinct set of VRN-A1 alleles compared to the diploids in this study, indicating an independent origin of spring tetraploid forms that likely occurred after combining of diploid genomes. These alleles were subsequently inherited by cultivated polyploid (tetraploid and hexaploid) descendants.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0473-x) contains supplementary material, which is available to authorized users.

The relationship between allelic variations ofVrn-1andPpd-1and agronomic traits in Korean wheat cultivars

The allelic variations at the Vrn-1 and Ppd-1 loci of Korean wheat cultivars were determined to evaluate their effects on agronomic traits and response to vernalization treatment. Korean wheat cultivars displayed high frequency of Vrn-D1a and Ppd-B1b alleles, but all of the tested cultivars carried vrn-A1, vrn-B1, Ppd-A1b and Ppd-D1a alleles at the corresponding loci. Regardless of vernalization treatments, Korean wheat cultivars carrying the vrn-D1 allele showed longer days to flag leaf unfolding than those with the Vrn-D1a allele. Unvernalized cultivars carrying the vrn-D1 allele also exhibited larger final leaf number than those with the Vrn-D1a allele. However, there were no significant differences in the response of the Korean wheat cultivars to vernalization treatment based on allelic variation at the Ppd-1 loci. With respect to agronomic traits, the Vrn-D1a allele was related to increased grain yield and decreased thousand kernel weight compared to the vrn-D1 allele. Cultivars carrying both Vrn-D1a and Ppd-B1b alleles produced higher grain yield (5.5 tons/ha) than those with the vrn-D1 and Ppd-B1b alleles (5.0 tons/ha).

Analysis of VRN1 gene in triticale and common wheat genetic background

In cereals, the transition from the vegetative stage to flowering is controlled in the main by the set of vernalization genes. Within these genes the most important role is played by VRN1, which encodes a MADS-box transcription factor, regulating the transition of shoot apical meristem to the reproductive phase. The level of vernalization requirement is strongly linked to the molecular structure of this gene. In this study we analyzed molecular mechanisms regulating the vernalization requirement in triticale on the basis of comparative analysis of the VRN1 locus between triticale (×Triticosecale Witt.) and common wheat (Triticum aestivum L.) genotypes. We also estimated the influence of VRN genotype on heading time and the winter hardiness of these two species. Molecular markers developed for VRN genotype detection in common wheat were successfully applied to an analysis of triticale genomic DNA. Subsequent analysis of the amplicons nucleotide sequence confirmed full similarity of the products obtained between triticale and common wheat. All winter triticale cultivars tested contained the recessive vrn-A1 allele, whereas all spring genotypes carried the dominant Vrn-A1a allele. Molecular analysis of the Vrn-B1 gene revealed the presence of the dominant Vrn-B1b allele in only one of the triticale genotypes analyzed (Legalo). The major system of determination of the vernalization requirement in triticale was transferred from common wheat without changes and is based on an alteration in the VRN1 gene promoter sequence within the A genome.

Copy number and haplotype variation at the VRN-A1 and central FR-A2 loci are associated with frost tolerance in hexaploid wheat.

The interaction between VRN - A1 and FR - A2 largely affect the frost tolerance of hexaploid wheat. Frost tolerance is critical for wheat survival during cold winters. Natural variation for this trait is mainly associated with allelic differences at the VERNALIZATION 1 (VRN1) and FROST RESISTANCE 2 (FR2) loci. VRN1 regulates the transition between vegetative and reproductive stages and FR2, a locus including several tandemly duplicated C-REPEAT BINDING FACTOR (CBF) transcription factors, regulates the expression of Cold-regulated genes. We identified sequence and copy number variation at these two loci among winter and spring wheat varieties and characterized their association with frost tolerance. We identified two FR-A2 haplotypes-'FR-A2-S' and 'FR-A2-T'-distinguished by two insertion/deletions and ten single nucleotide polymorphisms within the CBF-A12 and CBF-A15 genes. Increased copy number of CBF-A14 was frequently associated with the FR-A2-T haplotype and with higher CBF14 transcript levels in response to cold. Factorial ANOVAs revealed significant interactions between VRN1 and FR-A2 for frost tolerance in both winter and spring panels suggesting a crosstalk between vernalization and cold acclimation pathways. The model including these two loci and their interaction explained 32.0 and 20.7 % of the variation in frost tolerance in the winter and spring panels, respectively. The interaction was validated in a winter wheat F 4:5 population segregating for both genes. Increased VRN-A1 copy number was associated with improved frost tolerance among varieties carrying the FR-A2-T allele but not among those carrying the FR-A2-S allele. These results suggest that selection of varieties carrying the FR-A2-T allele and three copies of the recessive vrn-A1 allele would be a good strategy to improve frost tolerance in wheat.

Allele frequencies in the VRN-A1, VRN-B1 and VRN-D1 vernalization response and PPD-B1 and PPD-D1 photoperiod sensitivity genes, and their effects on heading in a diverse set of wheat cultivars (Triticum aestivum L.).

Heading of cereals is determined by complex genetic and environmental factors in which genes responsible for vernalization and photoperiod sensitivity play a decisive role. Our aim was to use diagnostic molecular markers to determine the main allele types in VRN-A1, VRN-B1, VRN-D1, PPD-B1 and PPD-D1 in a worldwide wheat collection of 683 genotypes and to investigate the effect of these alleles on heading in the field. The dominant VRN-A1, VRN-B1 and VRN-D1 alleles were present at a low frequency. The PPD-D1a photoperiod-insensitive allele was carried by 57 % of the cultivars and was most frequent in Asian and European cultivars. The PPD-B1 photoperiod-insensitive allele was carried by 22 % of the genotypes from Asia, America and Europe. Nine versions of the PPD-B1-insensitive allele were identified based on gene copy number and intercopy structure. The allele compositions in PPD-D1, PPD-B1 and VRN-D1 significantly influenced heading and together explained 37.5 % of the phenotypic variance. The role of gene model increased to 39.1 % when PPD-B1 intercopy structure was taken into account instead of overall PPD-B1 type (sensitive vs. insensitive). As a single component, PPD-D1 had the most important role (28.0 % of the phenotypic variance), followed by PPD-B1 (12.3 % for PPD-B1_overall, and 15.1 % for PPD-B1_intercopy) and VRN-D1 (2.2 %). Significant gene interactions were identified between the marker alleles within PPD-B1 and between VRN-D1 and the two PPD1 genes. The earliest heading genotypes were those with the photoperiod-insensitive allele in PPD-D1 and PPD-B1, and with the spring allele for VRN-D1 and the winter alleles for VRN-A1 and VRN-B1. This combination could only be detected in genotypes from Southern Europe and Asia. Late-heading genotypes had the sensitivity alleles for both PPD1 genes, regardless of the allelic composition of the VRN1 genes. There was a 10-day difference in heading between the earliest and latest groups under field conditions.Electronic supplementary materialThe online version of this article (doi:10.1007/s11032-014-0034-2) contains supplementary material, which is available to authorized users.

Molecular characterization of vernalization and response genes in bread wheat from the Yellow and Huai Valley of China

BackgroundFlowering time greatly influences the adaptation of wheat cultivars to diverse environmental conditions and is mainly controlled by vernalization and photoperiod genes. In wheat cultivars from the Yellow and Huai Valleys, which represent 60%-70% of the total wheat production in China, the large-scale genotyping of wheat germplasms has not yet been performed in terms of vernalization and photoperiod response alleles, limiting the use of Chinese wheat germplasms to a certain extent.ResultsIn this study, 173 winter wheat cultivars and 51 spring wheat cultivars from China were used to identify allelic variations of vernalization and photoperiod genes as well as copy number variations of Ppd-B1 and Vrn-A1. Two new co-dominant markers were developed in order to more precisely examine Vrn-A1b, Vrn-B1a, and Vrn-B1b. Two novel alleles at the Vrn-B3 locus were discovered and were designated Vrn-B3b and Vrn-B3c. Vrn-B3b had an 890-bp insertion in the promoter region of the recessive vrn-B3 allele, and Vrn-B3c allele had 2 deletions (a 20-bp deletion and a 4-bp deletion) in the promoter region of the dominant Vrn-B3a allele. Cultivar Hemai 26 lacked the Vrn-A1 gene. RT-PCR indicated that the 890-bp insertion in the Vrn-B3b allele significantly reduced the transcription of the Vrn-B3 gene. Cultivars Chadianhong with the Vrn-B3b allele and Hemai 26 with a Vrn-A1-null allele possessed relatively later heading and flowering times compared to those of Yanzhan 4110, which harbored recessive vrn-B3 and vrn-A1 alleles. Through identification of photoperiod genes, 2 new polymorphism combinations were found in 6 winter wheat cultivars and were designated Hapl-VII and Hapl-VIII, respectively. Distribution of the vernalization and photoperiod genes indicated that all recessive alleles at the 4 vernalization response loci, truncated “Chinese Spring” Ppd-B1 allele at Ppd-B1 locus and Hapl-I at the Ppd-D1 locus were predominant in Chinese winter wheat cultivars.ConclusionThis study illustrated the distribution of vernalization and photoperiod genes and identified 2 new Vrn-B3 alleles, 1 Vrn-A1-null allele, and two new Ppd-D1 polymorphism combinations, using developed functional markers. Results of this study have the potential to provide useful information for screening relatively superior wheat cultivars for better adaptability and maturity.

Molecular examination and genotype diversity of vernalization sensitivity and photoperiod response in old and modern bread wheat cultivars grown in Iran

Understanding the genetic factors governing developmental patterns and flowering time in breeding materials is required for the development of new wheat varieties for a specific environment. Iran is among the largest wheat producers in the arid and semi-arid regions of the Middle East and North Africa. The wheat germplasm grown in Iran is either developed nationally or is introduced from the CIMMYT global wheat program. For decades, the wheat breeding program in Iran focused on generating new varieties better able to grow in the predominant Iranian climatic conditions such as humidity at the reproductive stage, high temperature during reproductive stages (terminal heat stress), moderate temperature during the cropping season, and high probability of frost damage during early stages of growth. There have also been sub-programs aimed at developing drought and salinity-tolerant wheat cultivars in Iran. Knowledge of cultivars’ growth habits in Iran is currently limited to flowering in spring-sown nurseries. We identified allelic diversity in loci involved in vernalization response (Vrn) and photoperiod sensitivity (Ppd) in 60 bread wheat cultivars developed in Iran, CIMMYT, or ICARDA. This study revealed that the spring growth habit observed in most of the cultivars is conferred by a combination of recessive vrn-A1 and dominant Vrn-D1, Vrn-B1, and/or Vrn-B3 loci. This implies that most of the cultivars have minimal vernalization requirements for overwintering. Perhaps cold winters, even in the southern regions of Iran, provide sufficient vernalization conditions for cultivars possessing the recessive vrn-A1 allele. The germplasm investigated in this study revealed no evidence indicating selection for or against any specific Vrn and Ppd allele in our wheat breeding program.

Molecular genetical characterization of vernalization genesVrn-A1, Vrn-B1andVrn-D1in spring wheat germplasm from Russia and adjacent regions

We characterized a representative set of 42 spring wheat cultivars from Russia and adjacent regions for 3 Vrn loci. The 42 genotypes were screened, along with 3 genotypes of known Vrn genes, using previously published genome-specific polymerase chain reaction (PCR) 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 28 of 42 cultivars, whereas the promoter deletion allele Vrn-A1b was present in only 1 of the Russian cultivars (Triticum aestivum L. ‘Pyrothrix 28’). The intron deletion allele Vrn-A1c was not present in any tested cultivar. The dominant Vrn-D1 allele was found in 1 of the cultivars. Thirteen of the spring wheat cultivars tested here carry the recessive vrn-A1 allele. However, for 6 cultivars, there were inconsistencies between PCR data and genetic segregation analysis, showing the presence of the dominant Vrn-A1 gene. No inconsistencies were found in the case of Vrn-B1 locus. A new combination of specific primers allowed amplification of the common Vrn-B1a allele along with the novel Vrn-B1c allele, which was present in 17 of the studied cultivars (40%). Twenty-five cultivars (59%) had dominant alleles of Vrn-A1a and Vrn-B1 in combination. We showed the predominance of the Vrn-B1c allele among cultivars with monogenic control of vernalization in West Siberia and Kazakhstan. In the absence of epistatic effects of Vrn-A1, this allele causes an earlier heading time compared to Vrn-B1a, thereby avoiding early fall frosts. Suggestions are made concerning the origin and distribution of the Vrn-B1c allele among Russian spring wheats.

The combination of the Ppd and Vrn gene alleles determines the heading date in common wheat varieties

Combinations of the Ppd-D1, Vrn-A1, Vrn-B1, and Vrn-D1 gene alleles were analyzed using allele-specific primers in 56 varieties and lines of common wheat that were selected in the Lukyanenko Agricultural Research Institute. The significant influence of the combination of these alleles on the time of the beginning of the heading of the studied varieties was determined, which is manifested differently under conditions of long and short photoperiods, as well as depending on whether vernalization occurs. Molecular labeling of wheat Vrn and Ppd gene alleles allows breeders to consciously manipulate the original material (by having information in advance about the adaptive potential of the plants that are involved in the breeding process), as well as to select the allele combinations that are optimal for specific conditions of cultivation.

Genetics of flowering time in bread wheat Triticum aestivum: 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.

Copy Number Variation Affecting the Photoperiod-B1 and Vernalization-A1 Genes Is Associated with Altered Flowering Time in Wheat (Triticum aestivum)

The timing of flowering during the year is an important adaptive character affecting reproductive success in plants and is critical to crop yield. Flowering time has been extensively manipulated in crops such as wheat (Triticum aestivum L.) during domestication, and this enables them to grow productively in a wide range of environments. Several major genes controlling flowering time have been identified in wheat with mutant alleles having sequence changes such as insertions, deletions or point mutations. We investigated genetic variants in commercial varieties of wheat that regulate flowering by altering photoperiod response (Ppd-B1 alleles) or vernalization requirement (Vrn-A1 alleles) and for which no candidate mutation was found within the gene sequence. Genetic and genomic approaches showed that in both cases alleles conferring altered flowering time had an increased copy number of the gene and altered gene expression. Alleles with an increased copy number of Ppd-B1 confer an early flowering day neutral phenotype and have arisen independently at least twice. Plants with an increased copy number of Vrn-A1 have an increased requirement for vernalization so that longer periods of cold are required to potentiate flowering. The results suggest that copy number variation (CNV) plays a significant role in wheat adaptation.

A Novel Retrotransposon Inserted in the DominantVrn-B1Allele Confers Spring Growth Habit in Tetraploid Wheat (Triticum turgidumL.)

Vernalization genes determine winter/spring growth habit in temperate cereals and play important roles in plant development and environmental adaptation. In wheat (Triticum L. sp.), it was previously shown that allelic variation in the vernalization gene VRN1 was due to deletions or insertions either in the promoter or in the first intron. Here, we report a novel Vrn-B1 allele that has a retrotransposon in its promoter conferring spring growth habit. The VRN-B1 gene was mapped in a doubled haploid population that segregated for winter-spring growth habit but was derived from two spring tetraploid wheat genotypes, the durum wheat (T. turgidum subsp. durum) variety ‘Lebsock’ and T. turgidum subsp. carthlicum accession PI 94749. Genetic analysis revealed that Lebsock carried the dominant Vrn-A1 and recessive vrn-B1 alleles, whereas PI 94749 had the recessive vrn-A1 and dominant Vrn-B1 alleles. The Vrn-A1 allele in Lebsock was the same as the Vrn-A1c allele previously reported in hexaploid wheat. No differences existed between the vrn-B1 and Vrn-B1 alleles, except that a 5463-bp insertion was detected in the 5′-UTR region of the Vrn-B1 allele. This insertion was a novel retrotransposon (designated as retrotrans_VRN), which was flanked by a 5-bp target site duplication and contained primer binding site and polypurine tract motifs, a 325-bp long terminal repeat, and an open reading frame encoding 1231 amino acids. The insertion of retrotrans_VRN resulted in expression of Vrn-B1 without vernalization. Retrotrans_VRN is prevalent among T. turgidum subsp. carthlicum accessions, less prevalent among T. turgidum subsp. dicoccum accessions, and rarely found in other tetraploid wheat subspecies.

Contrasting cDNA-AFLP profiles between crown and leaf tissues of cold-acclimated wheat plants indicate differing regulatory circuitries for low temperature tolerance

Low-temperature (LT) tolerance in winter wheat (Triticum aestivum L.) is an economically important but complex trait. Four selected wheat genotypes, a winter hardy cultivar, Norstar, a tender spring cultivar, Manitou and two near-isogenic lines with Vrn-A1 (spring Norstar) and vrn-A1 (winter Manitou) alleles of Manitou and Norstar were cold-acclimated at 6°C and crown and leaf tissues were collected at 0, 2, 14, 21, 35, 42, 56 and 70days of cold acclimation. cDNA-AFLP profiling was used to determine temporal expression profiles of transcripts during cold-acclimation in crown and leaf tissues, separately to determine if LT regulatory circuitries in crown and leaf tissues could be delineated using this approach. Screening 64 primer combinations identified 4,074 and 2,757 differentially expressed transcript-derived fragments (TDFs) out of which 38 and 16% were up-regulated as compared to 3 and 6% that were down-regulated in crown and leaf tissues, respectively. DNA sequencing of TDFs revealed sequences common to both tissues including genes coding for DEAD-box RNA helicase, choline-phosphate cytidylyltransferase and delta-1-pyrroline carboxylate synthetase. TDF specific to crown tissues included genes coding for phospahtidylinositol kinase, auxin response factor protein and brassinosteroid insensitive 1-associated receptor kinase. In leaf, genes such as methylene tetrahydrofolate reductase, NADH-cytochrome b5 reductase and malate dehydrogenase were identified. However, 30 and 14% of the DNA sequences from the crown and leaf tissues, respectively, were hypothetical or unknown proteins. Cluster analysis of up-, down-regulated and unique TDFs, DNA sequence and real-time PCR validation, infer that mechanisms operating in crown and leaf tissue in response to LT are differently regulated and warrant further studies.

Diversity of Growth Habits and Their Association with VRN Allele of 81 American Wheat Lines

Eighty-one wheat accessions including 50 southern regional performance nursery (SRPN) lines and 31 northern regional performance nursery (NRPN) lines from the United States were tested to evaluate the growth habit by chilling treatments and to estimate the VRN allele variation with 19 pairs of published VRN primers. Two spring wheat accessions and 44 semi-spring wheat accessions were confirmed based on their chilling days' requirement and polymorphism was found at VRN loci. The Vrn-A1 allele had the highest frequency in the RPN accessions and VA1-CAPs markers identified growth habit of RPN lines. No polymorphism was found at the VRN3 loci and some polymorphism at the region of promoter and the first intron of VRN1 was not always consistent to growth habit in the wheat RPN accessions. The existence of variation in VRN alleles suggested that singly using the dominant Vr n allele is possible to extend the diversity of wheat accessions and improve their adaption to different environments in autumn-sowing region. This information will be useful for the cultivars exploitation and wheat breeding program.

Genetic regulation of developmental phases in winter wheat

The orderly development of winter wheat through its life cycle can be marked at three stages: stem elongation, heading date, and physiological maturity. The duration of a developmental phase between two stages is important in yield component generation. In this study the three developmental stages were characterized and 350 markers were mapped in a population of recombinant inbred lines (RILs) generated from a cross between two winter wheat cultivars (‘Jagger’ and ‘2174’). Three major QTLs were found to control variation in developmental process, and each of them was tightly associated with a known flowering gene, VRN-A1 on chromosome 5A, PPD-D1 on chromosome 2D, and VRN-D3 on chromosome 7D. The average contribution of the gene marker for each QTL to the total phenotypic variation (R2) was evaluated over 3 years. The effect of VRN-A1 ranged from 21.5% at stem elongation to 17.4% at physiological maturity. The effect of PPD-D1 was minor (6.7%) at stem elongation but increased to 29.7% at heading and 20.1% at physiological maturity. The effect of VRN-D3 was not detected at stem elongation but increased to 14.6% at heading and to 20.5% at physiological maturity. Hence, the VRN-A1 locus, the PPD-D1 locus, and the VRN-D3 locus had greatest impact on development at stem elongation, heading date, and physiological maturity, respectively. Whereas the Jagger VRN-A1 and VRN-D3 alleles accelerated development, the Jagger PPD-D1 allele delayed the developmental process due to its sensitivity to photoperiod. Our findings suggest that through the appropriate combination of alleles at these three loci one would be able to regulate the various developmental phases to accommodate different agricultural needs.

Allele Variation in Loci for Adaptive Response and Plant Height and its Effect on Grain Yield in Wheat

ABSTRACTIn the present study the microsatellite marker Xgwm261, linked to Rht 8 gene as well as allele specific markers for Ppd-D1 (Ppdl), Vrn-1A and Vrn-1D loci, and the gibberellic acid (GA) test were employed to examine the distribution of semi-dwarf and photoperiod response and vernalization genes in 21 old and modern Bulgarian, 4 introduced cultivars and 11 advanced breeding lines from the wheat germplasm collections of DAI, General Toshevo, Institute of Genetics, Sofia and IPGR, Sadovo. Several allele variants were identified at locus Xgwm261, among which the 192 bp allele can serve as a diagnostic marker for the presence of Rht 8 gene. The Ppd-D1a, vrn-A1 and vrn-D1 alleles were found in almost all wheat genotypes illustrating the relationship between the photoperiod and vernalization response and the adaptability to the regional environments. The photoperiod insensitivity allele (Ppd-D1a) was found in 93.3% of analyzed modern cultivars released during 1960–2003 and has been most probably introduced from the Ukrainian cultivar Bezostayal (192 bp allele at locus Xgwm261), which is the main donor of Rht 8 gene in Bulgarian wheat. All studied modern Bulgarian wheat cultivars excluding cv. Laska (vrn-A1, Vrn-D1) and cv. Gladiator 113 (Vrn-A1a, vrn-D1) were of winter type (vrn-A1 and vrn-D1). Allele Vrn-A1c determining also spring type was not found in the studied set of the old and modern Bulgarian wheat cultivars.The association between specific alleles at the Rht-B1, Xgwm261, Ppd-D1, Vrn1 loci with plant height, heading time and some yield related traits was also examined. The results suggested that allele combination Ppd-D1a/vrn-A1, vrn-D1 led to earlier heading than the allele combination Ppd-D1b/vrn-A1, vrn-D1.The recombinant genotypes obtained from the crosses between photoperiod sensitive spring and photoperiod insensitive winter type wheat behave differently under autumn and spring sowing. Under early spring sowing the earliest heading dates and yield per m2 were observed in lines with Ppd-D1a/Vrn-A1, vrn-D1 and Ppd-D1b/Vrn-A1, vrn-D1 allele combinations.

Allelic Variation at the VRN-1 Locus of Polish Cultivars of Common Wheat (Triticum aestivum L.)

At a molecular level, the length of the vernalization period of common wheat (Triticum aestivum L.) is determined mainly by three loci: VRN-1, VRN-2 and VRN-3. In hexaploid wheat, the Vrn-A1, Vrn-B1 and Vrn-D1 genes are dominant for spring growth habit and epistatic to the alleles for winter growth habit. We used DNA markers to determine the VRN-1 genotypes of 43 common wheat cultivars from the Polish register. All of the 30 examined winter wheat cultivars carried the recessive vrn-A1 allele, and all of the 13 analyzed spring cultivars carried the dominant Vrn-A1a allele. Moreover, 13 winter and 11 spring cultivars carried the dominant Vrn-B1 allele. These results confirmed that the hexaploid wheat growth habit is determined mainly by the VRN-A1 locus.

Vrn-D4 is a vernalization gene located on the centromeric region of chromosome 5D in hexaploid wheat

Natural variation in wheat requirement of long exposures to cold temperatures to accelerate flowering (vernalization) is mainly controlled by the Vrn-1, Vrn-2, Vrn-3, and Vrn-4 loci. The first three loci have been well characterized, but limited information is available for Vrn-4. So far, natural variation for Vrn-4 has been detected only in the D genome (Vrn-D4), and genetic stocks for this gene are available in Triple Dirk (TDF, hereafter). We detected heterogeneity in the Vrn-1 alleles present in different TDF stocks, which may explain inconsistencies among previous studies. A correct TDF seed stock from Japan carrying recessive vrn-A1, vrn-B1, and vrn-D1 alleles was crossed with three different winter cultivars to generate F(2) mapping populations. Most of the variation in flowering time in these three populations was controlled by a single locus, Vrn-D4, which was mapped within a 1.8 cM interval flanked by markers Xcfd78 and Xbarc205 in the centromeric region of chromosome 5D. A factorial ANOVA for heading time using Vrn-D4 alleles and vernalization as factors showed a significant interaction (P < 0.0001), which confirmed that the Vrn-D4 effect on flowering time is modulated by vernalization. Comparison of the different Triple Dirk stocks revealed that Vrn-B1, Vrn-D1, and Vrn-D4 all have a small residual response to vernalization, but Vrn-D4 differs from the other two in its response to short vernalization periods. The precise mapping and characterization of Vrn-D4 presented here represent a first step toward the positional cloning of this gene.