J. Sutka

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.

Genetic studies of frost resistance in wheat.

SummaryGenetic studies of frost resistance were performed on various wheat varieties using diallel, F2 monosomic and substitution analysis.A six-parental cross including reciprocals was carried out, and F1 hybrids and their parents were used for the freezing tests under controlled conditions. Both the general combining ability (GCA) and the specific combining ability (SCA) were significant, indicating additive and non-additive gene action in the inheritance of frost resistance. The high GCA∶SCA ratio revealed a preponderance of additive genetic variance. No significant reciprocal differences were found between the reciprocal crosses. The variance/covariance graphical analysis indicated the partial dominance of frost sensitivity. Frost sensitive varieties had the largest number of dominant genes, while frost resistant varieties had the highest proportion of recessive genes. The magnitude of the additive component of variation was higher than that of the dominance component, and the overall measure of the degree of dominance was smaller than one, so average dominance is incomplete. The increasing and decreasing alleles are not equally frequent at all loci. In this set of wheat varieties the values of narrow and broad heritability are relatively high.F2 monosomic analysis of the winter wheat variety ‘Arthur’ crossed with the monosomics of ‘Chinese Spring’ revealed that the average frost resistance of all the 21 monosomics was lower than that of the disomic. F2 monosomic hybrids 5A, 2B, 4B and 5D proved to be relatively frost resistant, while monosomics 3A, 3B and 6D were the most sensitive.The control of frost resistance in the set of chromosome substitution lines of the variety ‘Cheyenne’ into ‘Chinese Spring’ (with the exception of 2B) indicated that the genes responsible for the frost resistance of ‘Cheyenne’ are localised in chromosomes 5A, 7A, 4B, 5B, 4D and 5D.The genetic basis of frost resistance and problems of analysis are discussed.

Location of a gene for frost resistance on chromosome 5A of wheat

SummaryA gene for frost resistance on chromosome 5A of wheat was located using single chromosome recombinant lines from the cross between the substitution line Hobbit (Triticum spelta 5A) and Hobbit. In this sample of recombinant lines the locus for frost resistance, designated Fr1, is completely linked to the locus Vrn1 controlling vernalisation requirement. The results can be explained by a pleiotropic action of the Vrn1 locus or close genetic linkage between Vrn1 and Fr1. Further detailed study is necessary to resolve these alternative hypotheses.

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

Abstract 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.

Mapping genes for flowering time and frost tolerance in cereals using precise genetic stocks

The development of comprehensive genetic maps based on molecular markers has increased the power of genetical analysis immensely in the last few years. Characters previously recalcitrant to analysis, such as abiotic stress responses, are now amenable, and individual major genes and QTL mediating the variation can be identified. This has allowed the development of strategies for stress amelioration by adjusting the timing of the life cycle and introducing genes which enable the plant to tolerate stress. These strategies are illustrated here by showing how genes for vernalization response and cold tolerance on chromosomes 5A and 5D of wheat have been identified and located. Additionally, their relationships to genes in other species, such as barley and rice can be characterised through comparative mapping approaches, leading to strategies for their isolation using rice genomic tools.

Frost hardiness depending on carbohydrate changes during cold acclimation in wheat

The effect of cold hardening on the dynamics of frost tolerance and on carbohydrate metabolism was studied in the frost-sensitive Chinese Spring and the frost tolerant Cheyenne genotypes, and in some of the chromosome substitution lines, derived from the crosses of the donor Cheyenne to Chinese Spring. Total water-soluble carbohydrate, glucose, fructose, sucrose and fructan contents were measured in the leaves. Differences in the accumulation of carbohydrates associated with cold tolerance occurred early in response to low temperature. Total water-soluble carbohydrates and total fructan content increased continuously during the cold treatment in all genotypes, resulting in higher contents in tolerant genotypes than in sensitive ones. Their rate of accumulation correlated significantly with the frost tolerance after 19 days of cold treatment. During the cold acclimation, the maximum of fructose accumulation proceeded that of sucrose. Significant correlation was detected between fructose and sucrose content and frost hardiness on the 43rd day of cold treatment. Fructose accumulated to a greater extent in the most tolerant genotypes with a sharp peak on the 35th day of cold hardening, followed by a decrease. In the chromosome substitution lines, the considerable sucrose accumulation started after the 11th day with a maximum on the 43rd day of cold hardening, coinciding with the tolerance test.

Detection of wheat-barley translocations by genomic in situ hybridization in derivatives of hybrids multiplied in vitro

Wheat-barley translocations were identified by genomicin situ hybridization (GISH) in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) × barley (Hordeum vulgare L. cv. Betzes) hybrids. The regenerated hybrids were pollinated with the wheat line Martonvásári 9 kr1. Five translocated wheat-barley chromosomes were recovered among 51 BC2F2 progeny from the in vitro regenerated wheat × barley hybrids. All were single breakpoint translocations with the relative positions of the breakpoints ranging from the centromere to about 0.8 of the relative arm length. Of the four translocations with intercalary breakpoints, three were transfers of terminal barley segments to wheat chromosomes; one was a transfer of a terminal wheat segment to a barley chromosome. Because of the absence of diagnostic N-bands, the identity of three barley segments could not be determined; in one translocation the barley chromosome involved had a NOR so it must have been 5H or 6H, and the centric translocation was 4HS.2BL. Following selfing, homozygotes of four translocations were selected. The experiment suggests that in vitro culture conditions are conducive for major genome rearrangements in wheat-barley hybrids.

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

Abstract 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.

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

Abstract 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.

Genetic control of frost tolerance in wheat (Triticum aestivum L.)

SummaryThe frost tolerance of winter wheat is one component of winter hardiness. If seedlings are frost resistant, it means that they can survive the frost effect without any considerable damage. To study the genetic control of frost tolerance, an artificial freezing test was used. Frost tolerance is controlled by an additive-dominance system. The results of diallel analyses indicate the importance of both additive and non-additive gene action in the inheritance of this character. The dominant genes act in the direction of lower frost tolerance and the recessive genes in the direction of a higher level of frost tolerance. The results of monosomic and substitution analyses show that at least 10 of the 21 pairs of chromosomes are involved in the control of frost tolerance and winter hardiness. Chromosomes 5A and 5D have been implicated most frequently. The geneFr1 (Frost 1) was located on the long arm of chromosome 5A. Crosses between cultivars, chromosome manipulation and the induction of somaclonal variation may be suitable methods for broadening the gene pool for frost tolerance.