The effect of VRN1 genes on important agronomic traits in high‐yielding Canadian soft white spring wheat

For reproductive success, flowering time must synchronize with favourable environmental conditions. Vernalization genes play a major role in accelerating or delaying the time to flowering. We studied how different vernalization (VRN1) gene combinations alter days to flowering and maturity and consequently the effect on grain yield and other agronomic traits. The study focussed on the effect of the VRN1 gene series (Vrn‐A1, Vrn‐B1 and Vrn‐D1) and their combinations. The Vrn gene group Vrn‐A1a, Vrn‐B1, vrn‐D1 was the earliest to flower and mature, while Vrn‐A1b, Vrn‐B1, vrn‐D1 was the latest to flower. Spring wheat lines with vrn‐A1, Vrn‐B1, Vrn‐D1 were the highest yielding and matured at a similar time as those having vernalization genes Vrn‐A1a, Vrn‐B1 and Vrn‐D1. The findings of this study suggest that the presence of Vrn‐D1 has a direct or indirect role in producing higher grain yield. We therefore suggest the introduction of Vrn‐D1 allele into higher‐yielding classes within Canadian spring wheat germplasm.     

Evaluation of cold hardiness in two sets of near-isogenic lines of wheat (Triticum aestivum) with polymorphic vernalization alleles

In wheat, variation at the orthologus Vrn‐1 loci, located on each of the three genomes, A, B and D, is responsible for vernalization response. A dominant Vrn‐1a allele on any of the three wheat genomes results in spring habit and the presence of recessive Vrn‐1b alleles on all three genomes results in winter habit. Two sets of near‐isogenic lines (NILs) were evaluated for DNA polymorphisms at their Vrn‐A1, B1 and D1 loci and for cold hardiness. Two winter wheat cultivars, ‘Daws’ and ‘Wanser’ were used as recurrent parents and ‘Triple Dirk’ NILs were used as donor parents for orthologous Vrn‐1 alleles. The NILs were analysed using molecular markers specific for each allele. Only 26 of 32 ‘Daws’ NILs and 23 of 32 ‘Wanser’ NILs had a plant growth habit that corresponded to the marker genotype for the markers used. Freezing tests were conducted in growth chambers programmed to cool to ?21.5°C. Relative area under the death progress curve (AUDPC), with a maximum value of 100 was used as a measure of death due to freezing. The average relative AUDPC of the spring habit ‘Daws’Vrn‐A1a NILs was 86.15; significantly greater than the corresponding winter habit ‘Daws’Vrn‐A1b NILs (42.98). In contrast, all the ‘Daws’Vrn‐A1bVrn‐B1aVrn‐D1b and Vrn‐A1bVrn‐B1bVrn‐D1a NILs (spring habit) had relative AUDPC values equal to those of their ‘Daws’ sister genotypes with Vrn‐A1bVrn‐B1bVrn‐D1b NILs (winter habit). The average AUDPC of spring and winter habit ‘Wanser’ NILs differed at all three Vrn‐A1, Vrn‐B1 and Vrn‐D1 locus comparisons. We conclude that ‘Daws’ and ‘Wanser’ have different background genetic interactions with the Vrn‐1 loci influencing cold hardiness. The marker for Vrn‐A1 is diagnostic for growth habit and cold hardiness but there is no relationship between the Vrn‐B1 and Vrn‐D1 markers and the cold tolerance of the NILs used in this study.     

Effect of VRN‐1 and PPD‐D1 genes on heading time in European bread wheat cultivars

The variation of the vernalization (VRN‐1) and photoperiod (PPD‐1) genes offers opportunities to adjust heading time and to maximize yield in crop species. The effect of these genes on heading time was studied based on a set of 245 predominantly spring cultivars of bread wheat from the main eco‐geographical regions of Europe. The genotypes were screened using previously published diagnostic molecular markers for detecting the dominant or recessive alleles of the major VRN‐1 loci such as: VRN‐A1, VRN‐B1, VRN‐D1 as well as PPD‐D1. We found that 91% of spring wheat cultivars contain the photoperiod sensitive PPD‐D1b allele. Photoperiod insensitive PPD‐D1a allele has been found mainly in southern region of Europe. For this region the monogenic control of vernalization by VRN‐B1 or VRN‐D1 dominant alleles is common, whereas in the remaining part of Europe, the combination of photoperiod sensitive PPD‐D1b allele with dominant VRN‐A1, VRN‐B1 and recessive vrn‐D1 alleles represents the most frequent genotype. Also, we revealed a significantly later (5–8 days) heading of the monogenically dominant genotypes at VRN‐B1 as compared to the digenic VRN‐A1 VRN‐B1 genotypes.     

The influence of a spring habit gene, Vrn-D1, on heading time in wheat

The adaptability of wheat cultivars to environmental conditions is known to be associated with a vernalization requirement, that is, spring/winter habit. To clarify the genetic effect of the spring habit gene, Vrn‐D1, on heading time in the field, recombinant inbred lines (RILs) with or without the Vrn‐D1 gene were produced from F₂ plants of the cross between ‘Nanbukomugi’ and ‘Nishikazekomugi’, non‐carrier and carrier cultivars of this gene, respectively. Using growth chambers with a controlled temperature and photoperiod, three components of heading time, i.e. vernalization requirement, photoperiodic sensitivity and narrow‐sense earliness (earliness per se), were evaluated in each RIL. RILs with the Vrn‐D1 gene (E lines) showed greatly reduced vernalization requirements and slightly shorter narrow‐sense earliness than RILs without Vrn‐D1 (L lines), although no difference in photoperiodic sensitivity was observed between the two groups. RILs were planted at four different sites in Japan and examined for their heading time in the field. E lines headed significantly earlier than L lines at all locations, indicating that the earliness of E lines is stable in various environmental conditions. These results indicated that spring habit caused by Vrn‐D1 gene, as well as narrow‐sense earliness, was responsible for heading time in the field.     

Development of a kompetitive allele‐specific PCR marker for selection of the mutated Wx‐D1d allele in wheat breeding

Waxy (Wx) protein is a key enzyme for synthesis of amylose in endosperm. Amylose content in wheat grain influences the quality of end‐use products. Seven alleles have been described at the Wx‐D1 locus, but only two of them (Wx‐D1b, Wx‐D1e) were genotyped with codominant markers. The waxy wheat line K107Wx1 developed by treating ‘Kanto 107’ seeds with ethyl methanesulphonate carries the Wx‐D1d allele. However, no molecular basis supports this nomenclature. In the present study, DNA sequence analysis confirmed that a single nucleotide polymorphism in the sixth exon of Wx‐D1 changed tryptophan at position 301 into a termination codon. Based on this sequence variation, a PCR‐based KASP marker was developed to detect this point mutation using 68 BC₈F₁ plants and 297 BC₈F₂ lines derived from the cross ‘Ningmai 14’*9/K107Wx1. Combined with codominant markers for the Wx‐A1 and Wx‐B1 alleles, waxy and non‐waxy near‐isogenic lines were distinguished. The KASP marker was efficient in identifying the mutant allele and can be used to transfer waxiness to elite lines.     

Mapping of the Vrn-B1 gene in Triticum aestivum using microsatellite markers

An F₂ population segregating for the dominant gene Vrn‐B1 was developed from the cross of the substitution line ‘Diamant/'Miro‐novskaya 808 5A’ and the winter wheat cultivar ‘Bezostaya 1′. Microsatellite markers (Xgwm and Xbarc) with known map locations on chromosome 5B of common wheat were used for mapping the gene Vrn‐B1. Polymorphism between parental varieties was observed for 28 out of 34 microsatellite markers (82%). Applying the quantitative trait loci mapping approach, the target gene was mapped on the long arm of chromosome 5B, closely linked to Xgwm408. The map position of Vrn‐B1 suggests that the gene is homoeologous to other vernalization response genes located on the homoeologous group 5 chromosomes of wheat, rye and barley.     

Alien introgression of spring habit dominant genes into bread wheat

Summary Alien dominant genes of spring habit were introgressed into bread wheat. The introgression was undertaken by simple crossing of winter bread wheat to related spring species or genera, followed by backcrossing to winter bread wheat, and did not involve the use of the ph mutants or embryo culture. The introgressed genes were located mostly on chromosomes of homoeologous group 5, and were allelic to the known Vrn genes in bread wheat. Nevertheless three groups of lines were discovered with the genes possibly located on other chromosomes. These genes were non-allelic to each other and to known Vrn genes and were designated Vrn6 ^Sc , Vrn7 ^Sc (introgressed from Secale cereale) and Vrn8 ^Ts (from Triticum sphaerococcum).     

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

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

The effects on grain endosperm structure of an introgression from <Emphasis Type="Italic">Aegilops speltoides</Emphasis> Tausch. into chromosome 5A of bread wheat

The endosperm structure of the wheat kernel determines its end-use quality. The known diversity in endosperm structure is related to the Pina-D1 and Pinb-D1 genes comprising the Ha locus on chromosome 5DS. We studied the effect of a gene introduced into bread wheat from the diploid relative, Aegilops speltoides, a putative donor of the B genome. Grain hardness and vitreousness were investigated in lines with homoeologous introgressions into chromosome 5A of spring wheat cultivar ‘Rodina’. One introgression changed the endosperm texture from hard to soft and had the same effect when transferred to other wheat genotypes. This indicated that its action was analogous to the dominant allele at the Ha locus. The temporary symbol Ha–Sp is given to the gene. Segregation for vitreousness in F₃ offspring from monosomic hybrids was also investigated. Genetic variability for endosperm structure in wheat may be extended by manipulating both hardness and vitreousness. Wheat germplasm with introgressions from wild relatives can increase the genetic variability of milling characteristics.     

The influence of photoperiod on the Vrn-H2 locus (4H) which is a major determinant of plant development and reproductive fitness traits in a facultative × winter barley (Hordeum vulgare L.) mapping population

To determine the effect of Vrn‐H2 locus on plant developmental and agronomic traits, detailed controlled environment tests involving a factorial set of vernalization and photoperiod treatments were carried out using doubled haploid lines developed from a facultative (Vrn‐H2^?) × winter (Vrn‐H2⁺) barley cross. The allele phase at the Vrn‐H2 locus influenced heading date as well as the developmental and agronomic traits. The performance of Vrn‐H2⁺ lines was significantly influenced by vernalization: reproductive fitness traits showed significant decreases without vernalization. However, the effects of alleles at the Vrn‐H2 locus extended beyond simple satisfaction of the vernalization requirement. Vrn‐H2⁺ lines showed increased reproductive fitness compared with Vrn‐H2^? lines when vernalization was followed by a long photoperiod. The responses of the two Vrn‐H2 allele classes to photoperiod duration were quite different in terms of heading date, developmental and agronomic traits. These results suggest that alleles at the Vrn‐H2 locus – and/or tightly linked gene(s) – respond primarily to the exogenous signal of vernalization (temperature), but when the vernalization requirement has been fulfilled, they also respond to photoperiod duration.     

Identifying AFLP and microsatellite markers for vernalization response gene Vrn-B1 in hexaploid wheat using reciprocal mapping populations

Marker‐assisted selection may be useful for combining specific vernalization response (Vrn) alleles into a single wheat genotype for yield enhancement; however, DNA markers are only available for two of the three genes identified to date. The objectives of this study were to investigate reciprocal effects on days to heading using F₂ populations generated by cross‐hybridizing near‐isogenic lines (NILs) carrying spring (Vrn‐B1; TDB) and winter (vrn‐B1; TDC) alleles, and to identify markers linked to Vrn‐B1 through genetic linkage analysis. Heading data were recorded for 91 and 89 progeny from reciprocal mapping populations TDB/TDC and TDC/TDB, respectively, and significant (P < 0.0001) reciprocal and dominance effects were detected. Among 207 amplified fragment length polymorphisms primer pairs and seven wheat microsatellite markers screened, two and one, respectively, were linked distally to Vrn‐B1 on wheat chromosome 5BL. Microsatellite Xgwm408 was most closely linked to Vrn‐B1 at 3.9 and 1.1 cM in the TDB/TDC and TDC/TDB map, respectively. Reciprocal differences in recombination distances emphasize the importance of female parent choice when generating mapping populations. Molecular markers are now available for three Vrn loci in wheat.     

Diversity of five glutenin loci within durum wheat (Triticum turgidum L. ssp. durum (Desf.) Husn.) germplasm grown in Algeria

The HMW and B‐LMW glutenin subunits composition of 120 durum wheat germplasm grown in Algeria was examined using SDS‐PAGE. All together, 39 glutenin patterns were detected, including eight for HMW and 21 for B‐LMW glutenin subunits. Twenty‐six different alleles were identified for the five glutenin loci studied, that is, Glu‐A1 (3), Glu‐B1 (7), Glu‐A3 (5), Glu‐B3 (9) and Glu‐B2 (2). Two new alleles were found at Glu‐B3 locus: Glu‐B3new₁ encodes for five subunits (7 + 8 + 14 + 16 + 18) and Glu‐B3new₂ codes for five subunits (4 + 6* + 12 + 15 + 15*), of which subunit 15* with mobility between bands 15–16 was not described previously. At the Glu‐1 loci, the Glu‐A1c/Glu‐B1e allelic composition was predominant. For the B‐LMW glutenins, the most common allelic composition was Glu‐A3a/Glu‐B3a/Glu‐B2a. The collection analysed shows glutenin alleles and allele combinations related to high gluten strength. This information could be useful to select varieties with improved quality and also as a source of genes to develop new lines when breeding for quality.     

Survey and new insights in the application of PCR‐based molecular markers for identification of HMW‐GS at the Glu‐B1 locus in durum and bread wheat

Wheat, among all cereal grains, possesses unique characteristics conferred by gluten; in particular, high molecular weight glutenin subunits (HMW‐GS) are of considerable interest as they strictly relate to bread‐making quality and contribute to strengthening and stabilizing dough. Thus, the identification of allelic composition, in particular at the Glu‐B1 locus, is very important to wheat quality improvement. Several PCR‐based molecular markers to tag‐specific HMW glutenin genes encoding Bx and By subunits have been developed in recent years. This study provides a survey of the molecular markers developed for the HMW‐GS at the Glu‐B1 locus. In addition, a selection of molecular markers was tested on 31 durum and bread wheat cultivars containing the By8, By16, By9, Bx17, Bx6, Bx14 and Bx17 Glu‐B1 alleles, and a new assignation was defined for the ZSBy9_aF1/R3 molecular marker that was specific for the By20 allele. We believe the results constitute a practical guide for results that might be achieved by these molecular markers on populations and cultivars with high variability at the Glu‐B1 locus.     

Genotype analysis of the wheat semidwarf Rht‐B1b and Rht‐D1b ancestral lineage

In wheat, semidwarfism resulting from reduced height (Rht)‐B1b and Rht‐D1b was integral to the ‘green revolution’. The principal donors of these alleles are ‘Norin 10’, ‘Seu Seun 27’ and ‘Suwon 92’ that, according to historical records, inherited semidwarfism from the Japanese landrace ‘Daruma’. The objective of this study was to examine the origins of Rht‐B1b and Rht‐D1b by growing multiple seed bank sources of cultivars comprising the historical pedigrees of the principal donor lines and scoring Rht‐1 genotype and plant height. This revealed that ‘Norin 10’ and ‘Suwon 92’ sources contained Rht‐B1b and Rht‐D1b, but the ‘Seu Seun 27’ source did not contain a semidwarf allele. Neither Rht‐B1b nor Rht‐D1b could be definitively traced back to ‘Daruma’, and both ‘Daruma’ sources contained only Rht‐B1b. However, ‘Daruma’ remains the most likely donor of Rht‐B1b and Rht‐D1b. We suggest that the disparity between historical pedigrees and Rht‐1 genotypes occurs because the genetic make‐up of seed bank sources differs from that of the cultivars actually used in the pedigrees. Some evidence also suggests that an alternative Rht‐D1b donor may exist.     

Characterization of QEet.ocs-5A.1, a quantitative trait locus for ear emergence time on wheat chromosome 5AL

QEet.ocs‐5A.1, a quantitative trait locus controlling ear emergence time, has been detected on wheat chromosome 5AL using single chromosome recombinant lines (SCRs) developed from a cross between ‘Chinese Spring’ (CS) (‘Cappelle‐Desprez’ 5A) and CS (Triticum spelta 5A). This locus has little influence on grain yield and its components, and thus has breeding potential for changing ear emergence time without yield reduction. To characterize the phenotypic expression of QEet.ocs.1 and to test its interaction with the Vrn‐A1 gene for vernalization response, six near‐isogenic SCRs differing for these two gene regions were grown together with the parental controls under different vernalization and photoperiod regimes. The T. spelta allele of QEet.ocs.1 accelerated heading time when vernalization and photoperiod were satisfied, demonstrating that the function of this QTL is earliness per se. There was no interaction between Vrn‐A1 and QEet.ocs.1.     

Use of non‐adapted quantitative trait loci for increasing Fusarium head blight resistance for breeding semi‐dwarf wheat

Semi‐dwarf wheat is an important prerequisite for releasing a successful commercial cultivar in high‐yielding environments. In Northern Europe, this aim is achieved by using one of the dwarfing genes Rht‐B1 (formerly known as Rht‐1) or Rht‐D1 (Rht‐2). Both genes, however, result in a higher susceptibility to Fusarium head blight (FHB). We analysed the possibility to use the two non‐adapted FHB resistance quantitative trait loci Fhb1 and Fhb5 (syn. QFhs.ifa‐5A) to counterbalance the negative effect of the dwarfing allele Rht‐D1b in a winter wheat population of 585 doubled‐haploid (DH) lines segregating for the three loci. All lines were inoculated with Fusarium culmorum at four locations and analysed for FHB severity, plant height, and heading date. The DH population showed a significant (p < 0.001) genotypic variation for FHB severity ranging from 3.6% to 65.9% with a very high entry‐mean heritability of 0.95. The dwarfing allele Rht‐D1b reduced plant height by 24 cm, but nearly doubled the FHB susceptibility (24.74% vs. 12.74%). The resistance alleles of Fhb1 and Fhb5 reduced FHB susceptibility by 6.5 and 11.3 percentage points, respectively. Taken all three loci together, Fhb5 alone was already able to reduce FHB susceptibility to the same extent as Rht‐D1b increased it. This opens new avenues for selecting semi‐dwarf wheat by marker‐assisted introgression of Fhb5 without the enhancement of FHB susceptibility.     

Adaptation and ecological differentiation in wheat with special reference to geographical variation of growth habit and Vrn genotype

Geographical variation of growth habit was studied for 749 landraces from various parts of the world, with special reference to their adaptation and ecogeographical differentiation. The total frequency of spring‐type landraces was 49.9%, and varied between localities. Spring‐type landraces were frequent in two distinct areas where the average January temperature was either below ‐7°C or above 4°C, with winter‐type landraces in areas from ‐7°C to 4°C. These results indicated that geographical variation of growth habit is closely related to the degree of winter coldness. An analysis of the Vrn genotype for 216 spring‐type landraces demonstrated the uneven distribution of four Vrn genes, with Vrn4 being the least frequent. The adaptive Vrn genotype was different between localities. Genotypes carrying Vrn‐A1 and additional Vrn gene(s) were frequent in two distinct areas where the average January temperature was either below ‐7°C or over 10°C, while genotypes with any of three Vrn genes, except Vrn‐A1, adapted to areas with temperatures from 4°C to 10°C. Therefore, it was concluded that the adaptability of wheat landraces differed depending on their growth habit and Vrn genotype, and that ecotypes with different Vrn genotypes were allopatrically distributed as a result of adaptation to different winter temperature. However, the differential distribution of Vrn‐B1, Vrn‐D1 and Vrn4 could not be explained by their adaptability, and might reflect the polyphyletic origin of common wheat.     

The high and low molecular weight glutenin subunit polymorphism of Algerian durum wheat landraces and old cultivars

The high molecular weight (HMW) and B‐zone low molecular weight (B‐LMW) glutenin subunit composition of 45 Algerian durum wheat (Triticum turgidum L. var. durum) landraces and old cultivars were examined by sodium‐dodecyl‐sulphate polyacrylamide gel electrophoresis (SDS‐PAGE). Nine accessions were heterogeneous and presented two or three genotypes. All together, 33 glutenin patterns were detected, including 12 for HMW and 15 for B‐LMW glutenin subunits. Twenty‐four different alleles were identified for the five glutenin loci studied, Glu‐A1 (3), Glu‐B1 (6), Glu‐A3 (8), Glu‐B3 (5) and Glu‐B2 (2). Five new alleles were found, three at Glu‐A3 and two at Glu‐B3. At the Glu‐1 loci, the Glu‐A1c‐Glu‐B1e allelic composition was predominant (31%). For the B‐LMW glutenins, the most common allelic composition was Glu‐A3a‐Glu‐B3a‐Glu‐B2a (36%). The collection analysed shows a high percentage of glutenin alleles and allele combinations related to high gluten strength, together with some others that have not been tested so far. This information could be useful to select local varieties with improved quality and also as a source of genes to develop new lines when breeding for quality.     

Effects of the Gpc‐B1 locus on high grain protein content introgressed into Argentinean wheat germplasm

Wheat grain protein content (GPC) is important for human nutrition and has a strong influence on the quality of pasta and bread. The objective of this study was to analyse the introduction of the Gpc‐B1 allele into two Argentinean bread wheat cultivars. Near‐isogenic lines were developed in ‘ProINTA Oasis’ and ‘ProINTA Granar’ using marker‐assisted selection. Gpc‐B1 lines showed a significant (P = 0.01) increase in GPC and a significant (P = 0.001) decrease in grain weight in comparison with control lines without Gpc‐B1. Differences in yield were not significant (P = 0.49) between lines. Gpc‐B1 lines significantly reduced (P = 0.02) straw nitrogen concentration at maturity and significantly increased (P = 0.02) the nitrogen harvest index. When data were analysed by genotype and environment, differences in some analysed parameters were found, indicating that Gpc‐B1 expression may be affected by different genetic backgrounds and environmental conditions. These results suggest that the introgression of the Gpc‐B1 allele into Argentinean wheat germplasm could be a valuable resource for improving GPC with no detrimental effect on grain yield.