Effect of the 5R(5A) alien chromosome substitution on the growth habit and winter hardiness of wheat

The growth habit, ear emergence time, and frost tolerance of wheat/rye substitution lines have been studied in cultivars Rang and Mironovskaya Krupnozernaya whose chromosome 5A is substituted with chromosome 5R of Onkhoyskaya rye. Hybrid analysis has demonstrated that the spring habit of the recipient cultivars Rang and Mironovskaya Krupnozernaya is controlled by dominant gene Vrn-A1 located in chromosome 5A. Onokhoyskaya rye has a dominant gene for the spring habit (Sp1) located in chromosome 5R. It has been found that the resultant 5R(5A) alien-substitution lines have a winter type of development and ears do not emerge during summer in plants sown in spring. The change in growth habit has been shown to be related to the absence of the rye Spl gene expression in the substitution lines. The winter hardiness of winter 5R(5A) alien-substitution lines has been studied under the environmental conditions of Novosibirsk. Testing the lines in the first winter demonstrated that their winter survival is 20-27%. The possible presence of the frost resistance gene homeoallelic to the known genes Fr1 and Fr2 of the common wheat located on chromosomes 5A and 5D, respectively, is discussed.     

RFLP mapping of the vernalization (Vrn1) and frost resistance (Fr1) genes on chromosome 5A of wheat

A population of single chromosome recombinant lines was developed from the cross between a frost-sensitive, vernalization-insensitive substitution line, ‘Chinese Spring’ (Triticum spelta 5A) and a frost-tolerant, vernalization-sensitive line, ‘Chinese Spring’ (‘Cheyenne’ 5A), and used to map the genes Vrn1 and Fr1 controlling vernalization requirement and frost tolerance, respectively, relative to RFLP markers located on this chromosome. The Vrn1 and Fr1 loci were located closely linked on the distal portion of the long arm of 5AL, but contrary to previous observations, recombination between them was found. Three RFLP markers, Xpsr426, Xcdo504 and Xwg644 were tightly linked to both. The location of Vrn1 suggests that it is homoeologous to other spring habit genes in related species, particularly the Sh2 locus on chromosome 7 (5H) of barley and the Sp1 locus on chromosome 5R of rye.     

RFLP mapping of genes affecting plant height and growth habit in rye

Summary RFLP mapping of chromosome 5R in the F₃ generation of a rye (Secale cereale L.) cross segregating for gibberellic acid (GA₃)-insensitive dwarfness (Ct2/ct2) and spring growth habit (Sp1/sp1) identified RFLP loci close to each of these agronomically important genes. The level of RFLP in the segregating population was high, and thus allowed more than half of the RFLP loci to be mapped, despite partial homozygosity in the parental F₂ plant. Eight further loci were mapped in an unrelated F₂ rye population, and a further two were placed by inference from equivalent genetic maps of related wheat chromosomes, allowing a consensus map of rye chromosome 5R, consisting of 29 points and spanning 129 cM, to be constructed. The location of the ct2 dwarfing gene was shown to be separated from the segment of the primitive 4RL translocated to 5RL, and thus the gene is probably genetically unrelated to the major GA-insensitive Rht genes of wheat located on chromosome arms 4BS and 4DS. The map position of Sp1 is consistent both with those of wheat Vrn1 and Vrn3, present on chromosome arms 5AL and 5DL, respectively, and with barley Sh2 which is distally located on chromosome arm 7L (= 5HL).     

Nucleotide sequence and molecular analysis of the low temperature induced cereal gene, BLT4

Summary The nucleotide sequence and derived amino acid sequence of a cDNA clone (BLT4) for a low temperature induced barley gene were determined. This gene, together with a small family of related genes, was shown to reside on chromosome 3. The BLT4 clone has homology with genes in wheat and oats. Its expression was studied in oats and in barley doubled haploid lines segregating for spring/winter habit and for frost hardiness. These analyses show that elevated steady state levels of BLT4 mRNA are produced in shoot meristematic tissue after 3 days low positive temperature treatment. The low temperature response was found in all barley doubled haploid lines and was therefore not associated specifically with either the spring/winter habit or frost hardiness. Elevated levels of BLT4 mRNA were also seen in drought-stressed barley and it is likely that this is a gene encoding a low molecular weight protein that is responsive to dehydrative stresses, such as cold and drought.The EMBL accession number for BLT4 is X56547 H. vulgare cDNA     

Location of a gene regulating cold-induced carbohydrate production on chromosome 5A of wheat

 Major changes in osmotic potential during cold acclimation are due to changes in sugar concentration, and there is a good correlation between sugar content and frost tolerance. The objective of the present study was to localize a gene(s) responsible for carbohydrate accumulation during cold acclimation on chromosome 5A of wheat using recombinant lines developed from the cross between the substitution lines Chinese Spring (Cheyenne 5A) and CS(Triticum spelta 5A). Previously, major genes influencing frost resistance (Fr1) and vernalization requirement (Vrn1) had been localized on the long arm of that chromosome. The T. spelta 5A chromosome carrying the Fr1 (frost-sensitive) allele for frost tolerance and the Vrn1 (spring-habit) allele for vernalization requirement did not have a major effect on the sucrose and fructan contents in the Chinese Spring background. On the other hand, the presence of Cheyenne alleles for vernalization requirement, vrn1, and frost tolerance, fr1, significantly increased sugar concentrations. A recombinant line thought to exhibit recombination between the Vrn1 and Fr1 loci suggested that the gene regulating sucrose accumulation was closely associated with, or else represented a pleiotropic effect of, Vrn1, but was separable from the Fr1 locus. Received: 3 March 1997 / Accepted: 7 March 1997     

Genetical analysis of chromosome 5A of wheat and its influence on important agronomic characters

Summary Chromosome 5A of bread wheat, Triticum aestivum carries the major gene, Vrnl, which is one of the main determinants of the winter/spring growth habit polymorphism in this species. Genetical analysis of this chromosome has been carried out using single-chromosome recombinant lines to establish the pleiotropic effects of this locus and two other major genes, q determining ear morphology and bl determining the presence of awns, on important agronomic characters. The three major genes were located on the long arm of chromosome 5A with a gene order of: centromere -bl-q-Vrnl. Analysis of quantitative characters from a winter sowing revealed pleiotropic effects of Vrnl or the effects of closely linked loci on the characters plant height, tiller number and spikelet number. However effects on ear emergence time were not associated with Vrnl but with q as were effects on spikelet number and ear length. In addition a locus determining yield/plant was located between Vrnl and q. Independant loci determining height and ear length were apparent on the short arm of chromosome 5A. From a spring sowing, however, there was a large pleiotropic effect of Vrnl on ear emergence time, as well as the effects previously detected. In addition, associated with q were effects on plant height and grain size which were not expressed from the winter sowing.     

Comparative mapping of the wheat chromosome 5A Vrn-A1 region with rice and its relationship to QTL for flowering time

 The vernalization gene Vrn-A1 on chromosome 5A is the predominant gene determining the spring/winter habit difference in bread wheat. Vrn-A1 was physically mapped using a set of deletion lines which located it to the region of chromosome 5A flanked by deletion breakpoints 0.68 and 0.78. This interval was shown to be homoeologous to a region of rice chromosome 3 that contains the flowering-time QTL Hd-6, previously mapped in a Nipponbare×Kasalath cross, and FLTQ1, a novel QTL identified by analysis of 78 F₃ families derived from a cross of ‘IR20’ב63–83’. Possible relationships between Vrn-A1 and rice QTL are discussed. Analysis of the chromosome 5A deletion lines showed evidence for a second, more proximal flowering-time effect located between deletion breakpoints 0.56 and 0.64. The proximal part of chromosome 5A is homoeologous to rice chromosome 9, on which two QTL were detected in the ‘IR20ב63–83’ cross. The possible relationship between these effects is also discussed. Received: 23 December 1997 / Accepted: 12 January 1998     

Frost tolerance in winter wheat cultivars: different effects of chromosome 5A and association with microsatellite alleles

Frost tolerance of ten Bulgarian winter wheat (Triticum aestivum L.) cultivars (Milena, Pobeda, Sadovo-1, Enola, Kristal, Laska, Svilena, Russalka, No301 and Lozen) and five foreign cultivars (Mironovskaya 808, Bezostaya-1, Rannaya-12, Skorospelka-35 and Chinese Spring) was studied in two experimental seasons following natural cold acclimation and in one experiment carried out in controlled acclimation conditions. Considerable intercultivar variability in plant survival was observed after freezing at ?21 °C following sufficient cold acclimation, or at ?18 °C following insufficient or controlled acclimation. In seven cultivars, the effects of chromosome 5A on frost tolerance were investigated in their F₂ hybrids with chromosome 5A monosomic lines of cultivars with high, intermediate and low frost tolerance. The effects of chromosome 5A depended on the stress severity and the genetic background of the hybrids and varied even in cultivars of similar frost tolerance and vernalization requirements. Effects of other chromosomes besides 5A on frost tolerance were assumed. The analysis of six microsatellite loci located in the interval from centromere to Vrn-1 on of chromosomes 5AL, 5BL and 5DL showed that the major loci determining frost tolerance in Bulgarian winter wheats were Fr-A2 on chromosome 5AL, and, to a lesser extent, Fr-B1 on chromosome 5BL. A strong association of the 176 bp allele at locus wmc327 tightly linked to Fr-A2 with the elevated frost tolerance of cvs. Milena, Pobeda, Sadovo-1, Mironovskaya-808 and Bezostaya-1 was revealed. Relatively weaker association between frost tolerance and the presence of the 172 bp allele at locus Xgwm639 tightly linked to Fr-B1 was also observed.     

Chromosome composition of wheat-rye lines and the influence of rye chromosomes on disease resistance and agronomic traits

Identification of the chromosomal composition of common wheat lines with rye chromosomes was carried out using genomic in situ hybridization and 1RS- and 5P-specific PCR markers. It was demonstrated that wheat chromosomes 5A or 5D were substituted by rye chromosome 5R in the wheat-rye lines. It was established that one of the lines with complex disease resistance contained rye chromosome 5R and T1RS.1BL, while another line was found to contain, in addition to T1RS.1BL, a new Robertsonian translocation, T5AS.5RL. Substitution of the wheat chromosome 5A with the dominant Vrn-A1 gene for the Onokhoiskaya rye chromosome 5R led to lengthening of the germination-heading period or to a change in the type of development. A negative influence of T1RS.1BL on SDS sedimentation volume and grain hardness was demonstrated, along with a positive effect of the combination of T1RS.1BL and 5R(5D) substitution on grain protein content. Quantitative traits of the 5R(5A) and 5R(5D) substitution lines were at the level of recipient cultivars. A line with two translocations, T1RS.1BL + T5AS.5Rl, appeared to be more productive as compared to the line carrying T1RS.1BL in combination with the 5R(5D) substitution.     

The relationship between vernalization requirement and frost tolerance in substitution lines of wheat

Two sets of wheat (Triticum aestivum L.) substitution lines for the homoeologous group 5 chromosomes, 5A, 5B and 5D, carrying vernalization genes (Vrn-A1, Vrn-B1, Vrn-D1) were used to study the relationship between vernalization requirement and winter survival, with respect to the induction and maintenance of frost tolerance. Substitution lines carrying dominant Vrn loci substituted from the spring cultivars Zlatka (5A), Chinese Spring (5D) and the alternative cultivar eská Pesívka (5B) into three different winter wheat backgrounds, Vala, Koútka and Zdar, showed lower winter survival by 20, 36, and 41 % for substitutions of 5B, 5A and 5D, respectively, compared to the original winter cultivars. Reciprocal substitution lines between two winter cultivars Mironovskaya 808 and Bezostaya 1 carrying different recessive alleles, vrn-A1, vrn-B1, vrn-D1, did not exhibit a modified induction of frost tolerance, but the duration of good frost tolerance, as well as the ability to survive the whole winter, was changed. In accordance with the model suggesting that genes for vernalization act as a master switch regulating the duration of frost tolerance, substitutions of homoeologous group 5 chromosomes induced, at first, frost tolerance at a level equal to the parental cultivar, and then, relative to the different extent of saturation of vernalization requirement, they gradually lost both frost tolerance and their ability to re-induce significant frost tolerance with a drop in temperature following warm periods in the winter.     

Two loci on wheat chromosome 5A regulate the differential cold-dependent expression of the cor14b gene in frost-tolerant and frost-sensitive genotypes

Although cold acclimation in cereals involves the expression of many cold-regulated genes, genetic studies have shown that only very few chromosomal regions carry loci that play an important role in frost tolerance. To investigate the genetic relationship between frost tolerance and the expression of cold-regulated genes, the expression and regulation of the wheat homolog of the barley cold-regulated gene cor14b was studied at various temperatures in frost-sensitive and frost-tolerant wheat genotypes. At 18/15 °C (day/night temperatures) frost-tolerant plants accumulated cor14b mRNAs and expressed COR14b proteins, whereas the sensitive plants did not. This result indicates that the threshold temperature for induction of the wheat cor14b homolog is higher in frost-resistant plants, and allowed us to use this polymorphism in a mapping approach. Studies made with chromosome substitution lines showed that the polymorphism for the threshold induction temperature of the wheat cor14b homolog is controlled by a locus(i) located on chromosome 5A of wheat, while the cor14b gene was mapped in Triticum monococcum on the long arm of chromosome 2A^m. The analysis of single chromosome recombinant lines derived from a cross between Chinese Spring/Triticum spelta 5A and Chinese Spring/Cheyenne 5A identified two loci with additive effects that are involved in the genetic control of cor14b mRNA accumulation. The first locus was tightly linked to the marker psr911, while the second one was located between the marker Xpsr2021 and Frost resistance 1 (Fr1). Received: 20 July 1999 / Accepted: 15 November 1999     

Preferential elimination of chromosome 5R of rye in the progeny of 5R5D dimonosomics

Transmission of chromosome 5R of rye (Secale cereale L.) and chromosome 5D of common wheat (Triticum aestivum L.) through gametes of 5R5D dimonosomics (2n = 42, 20W″ + 5R′ + 5D′) was studied. Chromosome 5R was found to have lower competitiveness as compared to 5D. Gametes with the rye chromosome were two times less often involved in the formation of a progeny. The combined frequency of the karyotypes of wheat (5D5D) and wheat monosomics (5D) was 11.6-fold higher than the frequency of the karyotypes of substitution lines (5R5R) and monosomics for the rye chromosome (5R). The karyotypes of 10.38% of hybrid plants had aberrant 5R chromosomes with different translocations formed as a result of breakages in the centromere and in the proximal region of the long arm. Telocentrics for the short arm t5RS, i5RS isochromosomes, and chromosomes with a terminal deletion T5RS.5RL-del were identified. The absence of amplification of SSR markers mapped on 5RS and the detection of PCR products for a number of 5RL markers (including the genome-specific rye marker Xrms115) permitted nine plants carrying only the long arm of chromosome 5R to be revealed. Since t5RL telocentrics were not detected by the cytological analysis, the results obtained allow us to suggest the presence of small intercalary translocations of the long arm of chromosome 5R in chromosome 5D or in other wheat chromosomes.     

Development of PCR-based markers on chromosome 5H for assisted selection of frost-tolerant genotypes in barley

Frost tolerance is an important trait for barley breeding. Field selection for this trait is not always efficient since, especially in Southern Europe, severe winter frost occurs erratically. Recent advances of cloned genes and molecular markers in barley provide molecular breeders with the means to develop new, simple PCR-based molecular markers, which can be used to select frost-tolerant genotypes quickly without stress simulation. This paper reports the development of two STS markers derived from the RFLP probes WG644 and PSR637, chosen as they are located on the long arm of homoeologous group 5 chromosomes of Triticeae, known to harbour the most important loci for frost tolerance. The two STS markers were validated together with one selected RAPD marker, OPA17, by separating two sets of winter and spring barley genotypes with different levels of frost tolerance. The ability of the developed markers to select segregant frost-tolerant and frost-susceptible genotypes was then investigated in a population of doubled haploid lines derived from a cross between a highly tolerant ('Nure') and a susceptible ('Tremois') genotype. In this population only two markers, OPA17 and Psr637 demonstrated their efficiency in dividing the phenotypes according to the parental alleles. These two markers mapped on the long arm of chromosome 5H, tightly linked to two frost tolerance QTLs. Two polymorphic bands of the WG644 STS were mapped: the former on the long arm of chromosome 5H (Wg644c) and the latter (Wg644b) on the long arm of chromosome 2H.     

Plant regeneration and somaclonal variation from cultured immature embryos of sister lines of rye and triticale differing in their content of heterochromatin

Summary Plants were regenerated from cultured immature embryos of two pairs of sister lines of triticale (X Triticosecale) cvs Rosner and Drira and five sister lines of rye (Secale cereale). The triticale lines differ in heterochromatic content of a particular rye chromosome (6R or 7R), while the rye lines differ in only one heterochromatic band. Variation in morphogenetic response was present between the triticale cultivars and between the rye lines. One of the rye lines (7RL+ +) showed a distinctive superior response in terms of somatic embryogenesis. These findings are discussed in relation to factors affecting morphogenetic response and genetic stability in culture.     

Mapping the nucleolus organizer region,seed protein loci and isozyme loci on chromosome 1R in rye

Summary The nucleolus organizer region located on the short arm of chromosome 1R of rye consists of a large cluster of genes that code for ribosomal RNA (designated the Nor-R1 locus). The genes in the cluster are separated by spacer regions which can vary in length in different rye lines. Differences in the spacer regions were scored in two families of F₂ progeny. Segregation also occurred, in one or both of the families, at two seed protein loci and at two isozyme loci also located on chromosome 1R. The seed protein loci were identified as the Sec 1 locus controlling -secalins located on the short arm of chromosome 1R and the Sec 3 locus controlling high-molecular-weight secalins located on the long arm of 1R. The two isozyme loci were the Gpi-R1 locus controlling glucose-phosphate isomerase isozymes and the Pgd 2 locus controlling phosphogluconate dehydrogenase isozymes. The data indicated linkage between all five loci and map distances were calculated. The results indicate a gene order: Pgd 2 ... Sec 3 ... [centromere] ... Nor-R1 ... Gpi-R1 ... Sec 1. Evidence was obtained that rye possesses a minor 5S RNA locus (chromosome location unknown) in addition to the major 5S RNA locus previously shown to be located on the short arm of chromosome 1R.     

Genetic analysis of rye (Secale cereale L.) Genetics of male sterility of the G-type

Summary The genetics and relationships between the genes in rye located in the nucleus and cytoplasm of the male sterility of the G-type were investigated. A factor inducing male sterility was found in the cytoplasms or rye cv Schlägler alt and rye cv Norddeutscher Champagner. Monogenic inheritance was observed in linkage tests. Using primary trisomies of rye cv Esto, the nuclear gene ms1 was found to be located on chromosome 4R. Modifying genes, probably masked in normal cytoplasm but expressed in male-sterility-inducing cytoplasm together with gene ms1, were located on chromosomes 3R (ms2) and 6R (ms3). Mono-, di-, and trigenic inheritance types were found in backcross progenies of trisomies.     

Chromosomal location of alcohol dehydrogenase gene(s) in rye,using wheat-rye addition lines

Following disc electrophoresis on standard gels, rye seed extracts showed two bands (ADH-3 and 5) for alcohol dehydrogenase. The ADH-3 band was homologous to the ADH band observed in other diploid species of the Triticinae, and with the ADH-3 band of 4 × and 6 × wheat. It is proposed that the rye isoenzymes ADH-3 and 5 are governed respectively, by the genes Adh _R1 and Adh _R2. Using bread wheat (Holdfast) lines with disomic addition of individual rye (King II) chromosomes, we found that the ADH-5 band was associated with the addition of rye chromosome IV (after Riley), indicating thereby that Adh _R2 gene is located on this chromosome. The products of Adh _R1 and Adh _R2 do not form active heterodimers, among themselves, but do form active dimers with wheat ADH monomers. It is suggested that the use of chromosomal addition lines may provide a method for locating genes for those enzymes, where the rye and wheat isoenzymes are electrophoretically distinct.     

Physical mapping of the Vrn-A1 and Fr1 genes on chromosome 5A of wheat using deletion lines

 Homozygous deletion lines of wheat for 5AL, generated in the variety ‘Chinese Spring’, were tested for flowering time without vernalization and for frost resistance after cold hardening. It was found that the Vrn-A1 gene for vernalization requirement mapped between breakpoints 0.68 and 0.78, whilst the frost resistance gene Fr1 was flanked by deletion breakpoints 0.67 and 0.68. This confirms previous evidence that these genes are linked but are not the pleiotropic effect of a single gene. A comparison between the physical and genetic maps for Vrn-A1 and Fr1 shows that the linear order is identical. These results indicate that cytogenetically based physical maps of Vrn-A1 and Fr1 loci, together with genetic maps, could be useful in the further study of genome synteny and in elaborating a gene cloning strategy. Received: 16 November 1998 / Accepted: 28 November 1998     

Phylogenetic relationships among genotypes of worldwide collection of spring and winter ryes (Secale cereale L.) determined by RAPD-PCR markers

Genetic similarities among 20 spring and 22 winter accessions of agronomically different ryes from fourteen countries were estimated by employing random amplified polymorphic DNA (RAPD) techniques. Cluster analysis of genetic distance data showed that 42 genotypes were readily classifiable into two main groups: spring and winter groups. Within the spring group, cultivars fell into a North European and an American-Chinese group. Cultivars of winter rye fell into four groups: Northern European, Russian, American and Chinese lines. A UPGMA-dendrogram based on genetic distances of cultivars of rye within the winter and spring groups showed that the clusters corresponded well to their geographical locations. The results indicated that isolation has played an important role in the evolution of rye, and that temporal isolation has influenced the genetic diversity of rye more than geographical isolation. In this experiment, RAPD proved to be a rapid, reliable and practicable method of revealing polymorphisms in rye populations.