The detection of allelic variants at the recessive vrn loci of winter wheat

Substitution lines with reciprocal substitutions of chromosomes containing recessive alleles of the homoeologous group 5 chromosomeVrn genes between varieties of winter wheat with high vernalisation requirement (‘Mironovskaya 808’) and low vernalisation requirements (‘Bezostaya 1’) have been created. On this basis the genetic determination of vernalisation requirement was established. Substitution lines Mironovskaya 808 (Bezostaya 1 5A), Mironovskaya 808 (Bezostaya 1 5B), Mironovskaya 808 (Bezostaya 1 5D) and reciprocal substitution lines Bezostaya 1 (Mironovskaya 808 5A), Bezostaya 1 (Mironovskaya 808 5B) and Bezostaya 1 (Mironovskaya 808 5D) were grown under different durations of vernalisation (3, 4, 5, 6, 7 and 8 weeks) and their response was evaluated. Photoperiodic sensitivity of the original parental genotypes was also determined. Reciprocal substitution lines of the same chromosome that carries the same vrn allele responded differently to vernalisation deficit. Differences have been shown between all group 5 reciprocal substitutions. Lines carrying chromosomes 5A and 5D of Mironovskaya 808 had a high vernalisation requirement whereas lines carrying chromosome 5B of Bezostaya 1 (vrn2^B) had a low vernalisation requirement. The reciprocal lines had a reverse requirement. This explains the different vernalisation requirements of the original varieties: Mironovskaya 808 with a high vernalisation requirement carries two alleles (vrn1^M and vrn3^M) in its genotype that increase the vernalisation requirement, whereas Bezostaya 1 with a lower requirement for vernalisation contains only one such allele (vrn2^B). By combination of the alleles in the lines with the substitution of chromosome 5B carrying vrn2 allele that in both original genotypes work inversely to the other alleles, transgressive genotypes have been formed: genotype vrn1^M vrn2^B vrn3^M determines a higher vernalisation requirement than original variety Mironovskaya 808, and genotype vrn1^B vrn2^M vrn3^B determines a lower vernalisation requirement than the original Bezostaya 1. An incomplete vernalisation requirement prolonged the time to heading, with exponential dependence on the vernalisation deficit, or prevented heading altogether. The original varieties further differed in photoperiodic sensitivity (Mironovskaya 808 sensitive, Bezostaya 1 less sensitive) that also influenced the background of substitution lines. The impact of the background on the heading time showed itself by about one week difference between Mironovskaya 808 and Bezostaya 1 grown under 8 weeks vernalisation and normal photoperiod. The difference between the lines with Mironovskaya 808 background and the lines with Bezostaya 1 background was approximately the same and was not significantly changed in different vernalisation variants of the lines. This difference may be caused by different photoperiodic sensitivity of the original varieties, but also by other genes, such as genes of earliness per se. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Temperature as a component of the expression of developmental responses in wheat

Summary Studies were made of days to ear emergence under the constant temperatures of 9, 14, 19 and 25°C and 16 h photoperiod in three sets of wheat lines each possessing genotypes differing for developmental responses.Days to ear emergence in three near-isogenic lines of the wheat cultivar Triple Dirk, which differed for vernalization response, increased as the strength of the response increased. At the four temperatures Triple Dirk D (Vrn 1 vrn 2) was not significantly different from normal Triple Dirk (Vrn 1 Vrn 2) but Triple Dirk B (vrn 1 Vrn 2) was significantly (P=0.01) later than normal Triple Dirk at each temperature. This indicates that the vrn 1 allele confers stronger vernalization response than vrn 2 over the range of temperatures (9–24°C). However, Triple Dirk C (vrn 1 vrn 2) failed to head after 120 days at each temperature indicating strong interaction between vrn 1 and vrn 2 with each other (and possibly the Triple Dirk back-ground) to give a much stronger vernalization response than predictions from additivity of their individual effects.The second set comprised the four Chinese Spring/Thatcher chromosome substitution lines CS/T 3B, 6B, 7B and 5D, plus Chinese Spring and Thatcher, and were grown in the unvernalized condition. CS/T 5D was similar in days to ear emergence as Chinese Spring at all four temperatures but the other three lines were earlier to ear emergence, particularly as the temperature increased. Days to ear emergence was fastest at 14°C in all lines, except CS/T 3B, in which it decreased progressively from 9 to 24°C.The third set of Chinese Spring and Thatcher and the homoeologous group 2 chromosomes of Thatcher substituted in Chinese Spring, the group which is considered to be involved in the control of photoperiod sensitivity. The three substitution lines responded differently to temperature compared with Chinese Spring and with each other, with chromosome 2D being the least, and chromosome 2B the most, responsive to temperature.     

Waiting for fine times: genetics of flowering time in wheat

To maximise yield potential in any environment, wheat cultivars musthave an appropriate flowering time and life cycle duration which`fine-tunes' the life cycle to the target environment. This in turn, requiresa detailed knowledge of the genetical control of the key components of thelife cycle. This paper discusses our current knowledge of the geneticalcontrol of the three key groups of genes controlling life-cycle duration inwheat, namely those controlling vernalization response, photoperiodresponse and developmental rate (`earliness per se', Eps genes).It also discusses how our ability to carry out comparative mapping of thesegenes across Triticeae species, and particularly with barley, is indicatingnew target genes for discovery in wheat. Major genes controllingvernalization response, the Vrn-1 series, have now been located bothgenetically and physically on the long arms of the homoeologous group fivechromosomes. These genes are homoeologous to each other and to thevernalization genes on chromosomes 5H of barley and 5R of rye.Comparative analysis with barley also indicates that other series ofvernalization response genes may exit on chromosomes of homoeologousgroups 4 (4B, 4D, 5A) and 1. The major genes controlling photoperiodresponse in wheat, the Ppd-1 genes, are located on the homoeologousgroup 2 chromosomes, and are homoeologous to a gene on barleychromosome 2H. Mapping in barley also indicates a photoperiod responselocus on barley 1H and 6H, indicating that a homoeologous series shouldexist on wheat group 1 and 6 chromosomes. In wheat, only a few`earliness per se loci have been located, such as on chromosomes ofhomoeologous group 2. However, in barley, all chromosomes appear tocarry such loci, indicating that several series of loci that affectdevelopmental rate independent of environment remain to be discovered.Overall, comparative studies indicate that there are probably twenty-fiveloci controlling the duration of the life-cycle, Vrn, Ppd and Eps genes, that remain to be mapped in wheat. There are major gaps inour knowledge of the detailed physiological effects of genes discovered todate on the timing of the life cycle from different sowing dates. This isbeing addressed by studying the phenology of isogenic and deletion lines inboth field and controlled environmental conditions. This has indicated thatthe vernalization genes have major effects on the rate of primodiaproduction, whilst the photoperiod genes affect the timing of terminalspikelet production and stem elongation, and these effects interact withsowing date.     

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.     

Effects of barley chromosome on heading characters in wheat-barley chromosome addition lines

Heading time in cereals is a composite character determined by vernalization requirement, photoperiodic sensitivity and narrow-sense earliness. To study the effects of added barley chromosomes on the heading characters in wheat, two sets of wheat-barley chromosome addition lines, i.e., ‘Betzes’ barley chromosomes 2H to 7H added to ’Chinese Spring‘ wheat (CS-Be2H to CS-Be7H) and ‘New Golden’ barley chromosomes 5H and 6H added to ‘Shinchunaga’ wheat (Shi-NG5H, Shi-NG6H), were examined for their heading characters. All barley chromosomes except Be6H affected vernalization requirement and/or narrow-sense earliness in CS or Shi. Be5H chromosome also slightly increased the photoperiodic sensitivity of CS. Shi-NG5H addition line showed significantly decreased vernalization requirement in comparison with Shi, whereas CS-Be5H did not show any difference from CS. The F₁ hybrid of the cross, Shi-NG5H × CS-Be5H, exhibited the same level of vernalization insensitivity as the Shi-NG5H addition line, and plants with and without a vernalization requirement segregated in a 1 : 3 ratio in the F₂ generation. These observations, together with previous reports, suggest that the decreased vernalization requirement in the Shi-NG5H addition line was caused by the presence of a major dominant gene for spring habit, Sh₂, located on the NG5H barley chromosome. Furthermore, this study revealed that the Sh₂ gene in barley has a similar but weaker effect than the wheat vernalization insensitive gene, Vrn1, on the vernalization response in wheat. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparative mapping and its use for the genetic analysis of agronomic characters in wheat

Summary The advent of molecular marker systems has made it possible to develop comparative genetic maps of the genomes of related species in the Triticeae. These maps are being applied to locate and evaluate allelic and homoeoallelic variation for major genes and quantitative trait loci within wheat, and to establish the pleiotropic effects of genes. Additionally, the known locations of genes in related species can direct searches for homoeologous variation in wheat and thus facilitate the identification of new genes. Examples of such analyses include the validation of the effects of Vrn1 on chromosome 5A on flowering time in different crosses within wheat; the indication of pleiotropic effects for stress responses by the Fr1 locus on chromosome 5A; the detection of homoeologous variation for protein content on the homoeologous Group 5 chromosomes; and the detection of a new photoperiod response gene Ppd-H1 in barley from homoeology with Ppd2 of wheat.     

Molecular mapping of genes determining hairy leaf character in common wheat with respect to other species of the Triticeae

Two major genes controlling leaf pubescence were mapped on chromosomes 4BL (Hl1) and 7BS (Hl2 ^Aesp ) in wheat (Saratovskaya 29) and a wheat/Aegilops introgression line (102/00^I), respectively, together with quantitative trait loci (QTLs) determining hairiness of the leaf margin (QHl.ipk-4B, QHl.ipk-4D) and auricle (QPa.ipk-4B, QPa.ipk-4D) on the long arms of chromosomes 4B and 4D, respectively. The QTLs on chromosome 4D were contributed by a synthetic wheat and, therefore, originated from Aegilops tauschii. The homoeologous group 4 wheat/A. tauschii genes/QTLs detected in the present study were aligned with the barley pubescence genes Hln/Hsh and Hs _b and the hairy peduncle rye gene Hp1. The locus seems to be pleiotropically responsible for the pubescence of different plant organs in different species of the Triticeae. Another homoeologous series may be present on the short arms of the homoeologous group 7 chromosomes, based on the results of an allelic test cross between the Chinese local cultivar Hong-mang-mai carrying Hl2 and the wheat/Aegilops speltoides introgression line 102/00^I.     

Effects of homoeologous group 1 and 6 chromosomes of the Cappelle-Desprez (Bezostaya 1) substitution lines on aspects of bread-making quality of wheat

Summary The group 1 and 6 inter-varietal chromosome substitution lines of Cappelle-Desprez (Bezostaya 1) were intercrossed along with the donor and recipient varieties, Cappelle-Desprez and Bezostaya 1, to give 36 genetically different families. The analysis of the means of these families showed that variation in SDS-sedimentation volume fitted a predominantly additive model. There were no significant within or between chromosome interactions among the group 1 and 6 chromosomes. Nor was there any evidence for interactions between these chromosomes and those of the background. Significant dominance/within chromosome interactions amongst the background chromosomes were however detected. Some of the positive effects on SDS-sedimentation were associated with increased grain hardness. Chromosome effects on % grain protein were not correlated with SDS-sedimentation.     

Identifying the alien chromosomes in wheat – Leymus multicaulis derivatives using GISH and RFLP techniques

Genomic in situ hybridization (GISH) and restriction fragment length polymorphism (RFLP) were used to identify the Leymus multicaulis (XXNN, 2n = 28) chromosomes in wheat-L. muliticaulis derivatives. Fifteen lines containing L. multicaulis alien chromosomes or chromosomal fragments were identified. All alien chromosomes or fragments in these 15 lines were from the X genome and none were from the N genome. Eleven L. multicaulis disomic addition lines and four translocation-addition lines were identified with chromosome rearrangements among homoeologous groups 2, 3, 6 and 7. Only homoeologous group 1 lacked rearrangements in addition or translocation chromosomes. The results revealed that translocation in non-homoeologous chromosomes widely exists in the Triticeae and therefore it is necessary to identify the alien chromosomes (segments) in a wheat background using these combined techniques. During the course of the work, probe PSR112, was found to detect X genome addition lines involving L. multicaulischromosomes. This may prove to be a valuable probe for the identification of alien chromosomes in a wheat background. This revised version was published online in August 2006 with corrections to the Cover Date.     

Determination of the genetic variation in the effect of temperature and daylength on bolting of Brassica campestris L.

Summary Effects of temperature and daylength on the bolting of a number of B. campestris accessions were investigated both in the open and in the IVT phytotron. From the results it was concluded that low temperature and genotype are the predominant factors with respect to bolting. Daylength has rather limited influence. One cultivar bolted more than 12 weeks earlier at 10 and 14°C than at 18°C. On the other hand some cultivars did not require vernalization at all.At 14°C and short daylength, depending on genotype, the period of time between sowing and 50% bolting ranged from 8 to 30 weeks. At this temperature the stimulating effect of long day (24 h), depending again on the genotype, ranged from 0 to 4 weeks. The genotypes most resistant to bolting appeared to be the stubble turnips (B. campestris ssp. rapa).In terms of seed(ling) vernalization at 5°C the cold requirement ranged from 0 to more than 3 weeks, depending on the genotype. The required vernalization periods at 5°C can be used as a criterion in screening for resistance to bolting.     

Genetic control of esterase-6 isozymes in hexaploid wheat and rye

Summary The EST-6 leaf esterase phenotypes from euploid, nullisomic-tetrasomic and rye chromosome addition and substitution lines of common wheat were determined using polyacrylamide gel electrophoresis. Evidence is presented to demonstrate that Est-6 is a new set of genes, that are expressed in the leaf. The Est-6 gene set were clearly distinguished from the Est-5 genes which are expressed in the grain. The three homoeoallelic loci, Est-A6, Est-B6 and Est-D6, were located on chromosomes 3A, 3B and 3D. An Est-R6 gene was located on chromosome 6R is involved in rye. Some considerations concerning homoeology between homoeologous group 3 of wheat and the rye chromosome 6R are made.     

Occurrence of numerically unreduced (2n) gametes in Alstroemeria interspecific hybrids and their significance for sexual polyploidisation

The F1 hybrids of seven diploid Alstroemeria species (2n=2x=16) were investigated for the production of numerically unreduced (2n) gametes and their mode of origin. Based on a survey of 17 interspecific hybrid combinations,consisting of 119 genotypes, it was found that the F1 hybrids of Chilean-Brazilian species mostly produced first division restitution (FDR) 2n gametes. These F1 hybrids were self-pollinated in order to obtain F2 seeds, which was an indication that the F1 plants also produced 2neggs simultaneously. All the F2 progeny plants were typical allotetraploids, most of which formed 16 bivalents and a small proportion formed multivalents during metaphase I stages of meiosis. Through genomic in situ hybridisation (GISH) it was proved that multivalent formation in F2plants, derived from A. inodora ×A. pelegrina hybrid, was due to homoeologous recombination but not from reciprocal translocations. In order to test the segregation pattern of the recombinant chromosomes, an F3 population from one genotype, P6C49-6, was investigated. The recombinant chromosomes assorted independently from each other supporting the hypothesis that the segregation of chromosomes in ring quadrivalents did not behave like those in translocation heterozygotes. It was concluded that in allopolyploids of Alstroemeria,bilateral sexual polyploidisation could accomplish genetic recombination by both homoeologous crossing-over as well as through the assortment of chromosomes. This revised version was published online in July 2006 with corrections to the Cover Date.     

The effects of whole chromosome substitutions differing in alleles for hybrid dwarfing and photoperiodic sensitivity on tissue culture response (TCR) in wheat

Summary The wheat varieties Chinese Spring, Marquis and Thatcher and five intervarietal Chinese Spring substitution lines for chromosomes 2B (2 lines), 2D (2 lines) and 4A^*, differing from the recipient variety in alleles for hybrid dwarfing genes and/or the photoperiodic response genes were analysed for tissue culture response (TCR). It could be demonstrated that only chromosome 2B has a major effect on TCR. Allelic variation at the hybrid dwarfing loci seems to have no effect on tissue culture performance, even in the combination D1D2D3 which gives the grass dwarf phenotype. Also the allelic constitution at the Ppd loci, gave no indication for a direct major effect of those alleles, however there seems to exist genetical factors for TCR on the homoeologous group 2 chromosomes which may be closely linked to the Ppd loci.     

Validation of the two-gene epistatic model for vernalization response in a winter × spring barley cross

A two gene epistatic model in which a dominant “winter growth habit” allele at Vrn-H2 encodes a repressor with a corresponding binding site in a recessive vrn-H1 allele explains the vernalization response phenotypes in an array of barley germplasm. In order to validate the model genetically, we developed an F ₂ population (and F ₂-derived F ₃ families) from the cross of Hardy (winter) × Jubilant (spring). Using gene-specific primers, we determined the Vrn-H1 and Vrn-H2 allele architecture of each F ₂ plant and we measured the growth habit phenotype of each F ₂ plant via phenotyping of its F ₃ progeny under controlled environment conditions. We used a set of treatments involving plus/minus vernalization under long photoperiod and vernalization under short photoperiod. Alleles at the two loci showed expected patterns of segregation and independent assortment. Under long day conditions, the two Vrn genes were the primary determinants of heading date, regardless of the vernalization treatment. Under short photoperiod, the effects of these loci were not significant. There was incomplete dominance at Vrn-H1: heterozygotes were significantly later to head than Vrn-H1Vrn-H1 genotypes. Vrn-H2 genotypes were also significantly later to head, even when plants were vernalized. These results validate the two-gene epistatic model for vernalization response under long-day conditions. The results under short photoperiod, and the variance in flowering with vernalization, confirm that while the two Vrn genes are the primary determinants of vernalization response, they are part of a larger interactome that determines the timing of the vegetative to reproductive transition.     

The use of wheat aneuploids for the chromosomal assignment of microsatellite loci

Summary The chromosomal assignment of 64 PCR-amplified microsatellite loci and 29 additional fragments amplified by the same primer pairs is described for bread wheat (Triticum aestivum). The distribution over the different chromosomes and chromosome arms appears to be random. The highest proportion of microsatellite loci is found on the B genome, followed by the A and D genome. About half of the primer pairs amplified unique fragments, while the other half amplified additional fragments. 25% of the primer pairs, mostly designed to clones of a PstI-library, amplify fragments on homoeologous chromosomes. In some cases, more than one fragment on a single chromosome or fragments on non-homoeologous chromosomes occurred. The use of an automated DNA sequencer accounts for the accurate resolution of multiple fragments and enables to differentiate between fragments, amplified by a single primer pair, with size differences as small as two base pairs.     

Detection of an earliness per se quantitative trait locus in the proximal region of wheat chromosome 5AL

A homoeologous quantitative trait locus to that of eps5L on barley chromosome 5H was identified in a syntenic region of wheat chromosome 5A. Wheat single chromosome recombinant lines (SCRs) were developed from a cross between ‘Chinese Spring’(‘Cappelle-Desprez’ 5A) and ‘Chinese Spring’(Triticum spelta 5A), these were grown together with the parental controls under different vernalization and photoperiod regimes. The variation for ear emergence time accelerated heading induced by the T. spelta segment indicated an effect associated with the Xcdo412-Xbcd9 interval. Since no differences between the SCRs and controls in responses to vernalization and photoperiod treatments were detected, this effect was identified as an earliness per se gene, Q Eetocs-5 A.2, which may be homoeologous to the eps5L quantitative trait locus of barley. Xbcd926 has been found to be closely linked to the rice flowering time quantitative trait loci, QHd9a or FLTQ2, on chromosome 9, suggesting possible relationships among the quantitative trait loci across wheat, barley and rice genomes.     

Effects of loci determining photoperiod sensitivity (Ppd-H1) and vernalization response (Sh2) on agronomic traits in the 'Dicktoo'×'Morex' barley mapping population

The objectives of this research were to determine the individual and interaction effects of the Ppd-H1 and Sh2 loci on agronomic traits under short- and long-photoperiod regimes. Nineteen doubled haploid (DH) lines from the ‘Dicktoo’בMorex’ mapping population, which represented the four genotypes at the Ppd-H1 and Sh2 loci, were pheno-typed in controlled environment photoperiods. Both Ppd-H1 and Sh2 had significant effects on several agronomic traits, in addition to their role in determining first node appearance and flowering time. The magnitude of these effects depended on daylight. Under long-day conditions (18 h) Ppd-H1, and under short-day conditions (12 h) Sh2 was a significant determinant of most characters. The interactions between these two loci were significant for several characters, particularly for yield components, under both long- and short-photoperiod regimes. Under the long-day treatment, Ppd-H1 influenced plant height through the determination of node number. There was an epistatic association between the two loci for both 1000-kernel weight and tillering. The combination of photoperiod insensitivity and vernalization requirement caused a significant increase in tillering. This was paralleled by a decrease in 1000-kernel weight. Under the long-day treatment, neither Ppd-H1 nor Sh2 influenced plant yield. Under short-day conditions, the combination of photoperiod insensitivity and vernalization requirement had a pronounced negative effect on plant yield.     

Genetic Control of Vernalization, Day-length Response, and Earliness per se by Homoeologous Group-3 Chromosomes in Wheat

To identify homoeologous group-3 chromosomes that carry genes for vernalization, day-length responses, and earliness per se, a series of aneuploid lines (mono-somics and tetrasomics) and chromosome-substitution lines in ‘Chinese Spring’ (CS) were surveyed under different vernalization and day-length regimes in controlled environments. The results indicated that genes on all three chromosomes of group 3 can have striking effects on ear-emergence time. The replacement of CS 3B by its homologues in ‘Lutescens 62’ and ‘Cheyenne’ produced an increased insensitivity to vernalization, while 3B homologues from ‘Ceska Presivka’ gave CS a remarkable sensitivity to vernalization. This provided evidence for multiple allelism at a new Vrn locus on chromosome 3B. A negative association between gene dosage and day-length response was found in CS 3D which was thought to carry a gene for promoting insensitivity to day-length. The behaviour of CS monosomic 3A and CS (Timstein 3A), in reducing numbers of days to heading independently of environmental stimuli, suggested the presence of earliness per se genes on this chromosome.