U. Lohwasser

Molecular mapping of genomic regions associated with wheat seedling growth under osmotic stress

A quantitative trait loci (QTL) approach was applied to dissect the genetic control of the common wheat seedling response to osmotic stress. A set of 114 recombinant inbred lines was subjected to osmotic stress from the onset of germination to the 8th day of seedling development, induced by the presence of 12 % polyethylene glycol. Root, coleoptile and shoot length, and root/shoot length ratio were compared under stress and control conditions. In all, 35 QTL mapping to ten chromosomes, were identified. Sixteen QTL were detected in controls, 17 under stressed conditions, and two tolerance index QTL were determined. The majority of the QTL were not stress-specific. In regions on five chromosome arms (1AS, 1BL, 2DS, 5BL and 6BL) the QTL identified under stress co-mapped with QTL affecting the same trait in controls, and these were classified as seedling vigour QTL, in addition to those expressed in controls. Tolerance-related QTL were detected on four chromosome arms. A broad region on chromosome 1AL, including five QTL, with a major impact of the gene Glu-A1 (LOD 3.93) and marker locus Xksuh9d (LOD 2.91), positively affected root length under stress and tolerance index for root length, respectively. A major QTL (LOD 3.60), associated with marker locus Xcdo456a (distal part of chromosome arm 2BS) determined a tolerance index for shoot length. Three minor QTL (LOD < 3.0) for root length and root/shoot length ratio under osmotic stress were identified in the distal parts of chromosome arms 6DL (marker locus Xksud27a) and 7DL (marker locus Xksue3b). Selecting for the favourable alleles at marker loci associated with the detected QTL for growth traits may represent an efficient approach to enhance the plants’ ability to maintain the growth of roots, coleoptile and shoots in drought-prone soils at the critical early developmental stages.

Microsatellite mapping of complementary genes for purple grain colour in bread wheat (Triticum aestivum) L.

Complementary genes for purple grain colour Pp1, Pp2, Pp3 (now designated Pp1, Pp3b, Pp3a, respectively) were mapped using crosses between purple-grained hexaploid wheats ‘Purple Feed’ – Pp1Pp1/Pp2Pp2 (Pp1Pp1/Pp3bPp3b), ‘Purple’ – Pp1Pp1/Pp3Pp3 (Pp1Pp1/Pp3aPp3a) with non-purple-grained cultivars ‘Novosibirskaya 67’ (‘N67’) and ‘Saratovskaya 29’ (‘S29’). The genes Pp2 (Pp3b) and Pp3 (Pp3a) were inherited as monofactorial dominant when purple-grained wheats were crossed to ‘N67’. Both were mapped in the centromeric region of the chromosome 2A. Therefore, they were suggested being different alleles at the same locus and designated Pp3a and Pp3b. In the crosses between purple-grained wheats and ‘S29’ a segregation ratio of 9 (purple) to 7 (non purple) was obtained suggesting a complementary interaction of two dominant genes, Pp1 and Pp3. To map Pp1 as a single gene, the influence of the other Pp gene was taken into consideration by determining the Pp3 genotype of the F2 plants. The gene was mapped on chromosome 7BL, about 24 cM distal to the centromere. The Pp1gene was shown to be non allelic to the Rc-1 (red coleoptile) and Pc (purple culm) genes, contrary to what was previously suggested. The colouration caused by the Pp genes has no effect on pre-harvest sprouting.

QTL mapping of the domestication traits pre-harvest sprouting and dormancy in wheat (Triticum aestivum L.)

A set of 75 recombinant inbred lines (RILs) of the ITMI mapping population was grown under field conditions in Gatersleben. The lines were evaluated for the domestication traits pre-harvest sprouting and dormancy (germinability). Main QTLs could be localized for pre-harvest sprouting on chromosome 4AL and dormancy on chromosome 3AL. In addition, 85 Triticum aestivum cv. “Chinese Spring”-Aegilops tauschii introgression lines grown under greenhouse conditions were researched. No QTL could be found for pre-harvest sprouting but a major QTL could be detected for dormancy on chromosome 6DL.

Investigation of antioxidant and rosmarinic acid variation in the sage collection of the genebank in Gatersleben.

The total phenolic and flavonoid contents and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reduction antioxidant power (FRAP) antioxidant capacities of 19 accessions of Salvia officinalis from the sage collection of the genebank in Gatersleben (Germany) were evaluated. The major phenolic compounds of sage, that is, rosmarinic acid, caffeic acid, carnosol, and carnosic acid, were quantified by reverse-phase high-performance liquid chromatography. The aerial parts of different individual plants of each accession were collected in two consecutive years from the same experimental field at the beginning of their flowering period. The results demonstrated a high variability between accessions. A general decreasing tendency from 2007 to 2008 was observed in most of the estimated parameters, that is, total phenolic, total flavonoid, rosmarinic acid, and caffeic acid contents and DPPH antioxidant activity. A slight opposite trend was obtained with the FRAP antioxidant capacity. Low but variable quantities of carnosol and carnosic acid were evaluated in the sample extracts. Individual plants within accessions were identified with high phenolic content and strong antioxidant activity. The rosmarinic acid content showed up to 8-fold differences between the lowest and the highest values. Overall, the study demonstrated a high variability in secondary metabolites present in sage, which could be used for breeding of highly antioxidative genotypes of S. officinalis.

Genetic studies of seed longevity in hexaploid wheat using segregation and association mapping approaches

Genebanks are entrusted with the storage, preservation and distribution of crop germplasm. Seed longevity is an important character in this context, but little is known regarding its genetic basis, largely because it is so strongly influenced by non-genetic factors. Here we present the outcome of a genetic dissection of seed longevity in bread wheat. We applied both a standard quantitative trait locus analysis based on segregation from a biparental cross, and an association analysis using a germplasm panel to detect marker trait associations. The latter revealed more loci than the former. Some of the genomic regions identified are known to contain genes determining spike architecture or aspects of biotic and abiotic stress responses. The results open perspectives for identification of favourable longevity alleles and the more accurate prediction of seed longevity in cereal germplasm collections.

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 (Hl2Aesp) in wheat (Saratovskaya 29) and a wheat/Aegilops introgression line (102/00I), 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 Hsb 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 102/00I.

Molecular mapping of quantitative trait loci (QTLs) controlling aluminium tolerance in bread wheat

Aluminium (Al) toxicity is a major constraint to crop productivity in acidic soils. A quantitative trait locus (QTL) analysis was performed to identify the genetic basis of Al tolerance in the wheat cultivar ‘Chinese Spring’. A nutrient solution culture approach was undertaken with the root tolerance index (RTI) and hematoxylin staining method as parameters to assess the Al tolerance. Using a set of D genome introgression lines, a major Al tolerance QTL was located on chromosome arm 4DL, explaining 31% of the phenotypic variance present in the population. A doubled haploid population was used to map a second major Al tolerance QTL to chromosome arm 3BL. This major QTL (QaltCS.ipk-3B) in ‘Chinese Spring’ accounted for 49% of the phenotypic variation. Linkage of this latter QTL to SSR markers opens the possibility to apply marker-assisted selection (MAS) and pyramiding of this new QTL to improve the Al tolerance of wheat cultivars in breeding programmes.

An association mapping analysis of dormancy and pre-harvest sprouting in wheat

Seed dormancy and pre-harvest sprouting are important traits in bread wheat. Bi-parental populations have permitted the identification of several genes/quantitative trait loci controlling these traits, mapping to various bread wheat chromosomes. Here, we report the use of association mapping to uncover the genetic basis of both traits in a panel of 96 diverse winter wheat cultivars to establish the presence of marker-trait associations on many chromosomes. Potential candidate genes were identified by studying the gene content of the chromosome bins into which the major marker trait associations mapped.

QTL analysis for thousand-grain weight under terminal drought stress in bread wheat ( Triticum aestivum L.)

Grain yield under post-anthesis drought stress is one of the most complex traits, which is inherited quantitatively. The present study was conducted to identify genes determining post-anthesis drought stress tolerance in bread wheat through Quantitative Trait Loci (QTLs) analysis. Two cultivated bread wheat accessions were selected as parental lines. Population phenotyping was carried out on 133 F2:3 families. Two field experiments and two experiments in the greenhouse were conducted at IPK-Gatersleben, Germany with control and post-anthesis stress conditions in each experiment. Thousand-grain weight was recorded as the main wheat yield component, which is reduced by post-anthesis drought stress. Chemical desiccation was applied in three experiments as simulator of post-anthesis drought stress whereas water stress was applied in one greenhouse experiment. Analysis of variance showed significant differences among the F2:3 families. The molecular genetic linkage map including 293 marker loci associated to 19 wheat chromosomes was applied for QTL analysis. The present study revealed four and six QTLs for thousand-grain weight under control and stress conditions, respectively. Only one QTL on chromosome 4BL was common for both conditions. Five QTLs on chromosomes 1AL, 4AL, 7AS, and 7DS were found to be specific to the stress condition. Both parents contributed alleles for drought tolerance. Taking the known reciprocal translocation of chromosomes 4AL/7BS into account, the importance of the short arms of homoeologous group 7 is confirmed for drought stress.

Discovery of loci determining pre-harvest sprouting and dormancy in wheat and barley applying segregation and association mapping

Three wheat and two barley populations were studied in order to find loci responsible for dormancy and pre-harvest sprouting. A classical quantitative trait loci analysis was combined with an association mapping approach. Many quantitative trait loci and marker trait associations could be detected on all seven chromosome groups of wheat and on the chromosomes 2H, 3H, 5H, 6H, and 7H of barley. Especially, the known regions on chromosomes 3A and 4A for wheat and 5H for barley were confirmed. Putative functions could be found via a candidate homologues search and via expressed sequence tag annotation. On chromosome 3A, the viviparous1 gene is located which is associated to preharvest sprouting and dormancy. On chromosome 4A, a protein is detected which belongs to the aquaporin family. In barley, an association with the aleurain gene on chromosome 5H was found. The expression of aleurain is regulated by abscisic acid and gibberelic acid. An influence of both hormones on dormancy and pre-harvest sprouting is known. It can be concluded that dormancy and pre-harvest sprouting are very complex traits regulated by multigenes and/or quantitative trait loci.