Axel Diederichsen

Genetic diversity within a range of cultivars and landraces of flax (Linum usitatissimum L.) as revealed by RAPDs

Analysis of the extent and distribution of genetic diversity incrop plants is essential for optimizing sampling and breedingstrategies. We used random amplified polymorphic DNA (RAPD)markers to assess genetic diversity and relationships in 22 Canadiancultivars, 29 selected world cultivars and 10 landraces of flax(Linum usitatissimum L.). RAPDvariation was generally low and more variation was detected among,than within, the investigated flax accessions. Based on 53 variableRAPD loci observed for the 61 accessions, the landraces had a lowerproportion of fixed recessive RAPD loci (0.427) (i.e.,more genetic variation) than all of the flax cultivars examined(0.492). The linseed cultivars had a lower proportion ofrecessive loci (0.469) than the fiber flax cultivars(0.529). Canadian linseed cultivars had a lower proportionof recessive loci (0.465) than the selected world flaxcultivars (0.512). A trend was also observed that the rateof loss in genetic variation in Canadian flax breeding programs overthe last fifty years was approximately two variable loci per 100 lociper 10 years. Clustering analyses based on similarity estimatesshowed that the fiber cultivars were more related (or similar toeach other) and were classified as a homogeneous group. All ofthe linseed cultivars were clustered in diverse groups with the ninelandrace accessions. Implications of these findings for flax breedingand germplasm management are discussed.

RAPD analysis of genetic relationships of seven flax species in the genus Linum L

The wild progenitor of the cultivated flax (Linumusitatissimum L.) has been long hypothesized to beL. angustifolium Huds., largely fromseveral phytogeographic cytogenetic and phenotypic studies, but no molecularstudies on the issue are found. In this study, we genotyped 12 flax accessionsrepresenting seven flax species in the genus Linum with 527RAPD loci from 29 informative RAPD primers and analyzed their geneticrelationships with simple matching, Dices and Jaccards similaritycoefficients. Large RAPD variations were found among the flax species.L. usitatissimum andL. angustifolium had a higher RAPDsimilarity than the other pairs of flax species and these two species wereconsistently clustered in the same group with all of the similarity coefficientsused. This molecular finding provides an additional support for the hypothesisof L. angustifolium as the wildprogenitor of cultivated flax.

Genetic characterization of a core collection of flax (Linum usitatissimum L.) suitable for association mapping studies and evidence of divergent selection between fiber and linseed types

BackgroundFlax is valued for its fiber, seed oil and nutraceuticals. Recently, the fiber industry has invested in the development of products made from linseed stems, making it a dual purpose crop. Simultaneous targeting of genomic regions controlling stem fiber and seed quality traits could enable the development of dual purpose cultivars. However, the genetic diversity, population structure and linkage disequilibrium (LD) patterns necessary for association mapping (AM) have not yet been assessed in flax because genomic resources have only recently been developed. We characterized 407 globally distributed flax accessions using 448 microsatellite markers. The data was analyzed to assess the suitability of this core collection for AM. Genomic scans to identify candidate genes selected during the divergent breeding process of fiber flax and linseed were conducted using the whole genome shotgun sequence of flax.ResultsCombined genetic structure analysis assigned all accessions to two major groups with six sub-groups. Population differentiation was weak between the major groups (FST = 0.094) and for most of the pairwise comparisons among sub-groups. The molecular coancestry analysis indicated weak relatedness (mean = 0.287) for most individual pairs. Abundant genetic diversity was observed in the total panel (5.32 alleles per locus), and some sub-groups showed a high proportion of private alleles. The average genome-wide LD (r2) was 0.036, with a relatively fast decay of 1.5 cM. Genomic scans between fiber flax and linseed identified candidate genes involved in cell-wall biogenesis/modification, xylem identity and fatty acid biosynthesis congruent with genes previously identified in flax and other plant species.ConclusionsBased on the abundant genetic diversity, weak population structure and relatedness and relatively fast LD decay, we concluded that this core collection is suitable for AM studies targeting multiple agronomic and quality traits aiming at the improvement of flax as a true dual purpose crop. Our genomic scans provide the first insights into candidate regions affected by divergent selection in flax. In combination with AM, genomic scans have the ability to increase the power to detect loci influencing complex traits.

Phenotypic and Molecular (RAPD) Differentiation of Four Infraspecific Groups of Cultivated Flax ( Linum usitatissimum L. subsp. usitatissimum )

Based on agro-botanical characterization data, 3101 accessions of cultivated flax (Linum usitatissimum L. subsp. usitatissimum) from the flax collection held by Plant Gene Resources of Canada (PGRC) were grouped into four infraspecific groups according to the classification proposed by Kulpa and Danert (1962) . The objective of this study was to investigate phenotypic and RAPD variation within and among the four groups to better understand phenotypic and genotypic differentiation within the genepool of cultivated flax. The results of the phenotypic characterization of characters defining the convarieties (capsule dehiscence, plant height, technical stem length and 1000 seed weight) and of other quantitative (petal width, oil content in seeds) and qualitative (RAPD, petal colour, anther colour, petal longitudinal folding and margin folding, ciliation of capsule septa, seed colour) are presented using descriptive statistics. The most frequent convariety in the PGRC genebank was intermediate flax (convar. usitatissimum; 80.7 %), followed by fibre flax (convar. elongatum Vav. et Ell. in Wulff; 13.4%), large-seeded flax (convar. mediterraneum [Vav. ex Ell.] Kulpa et Danert; 5.6%) and dehiscent flax (convar. crepitans [Boenningh.] Kulpa et Danert; 0.3%). Analyses of RAPD data and two qualitative characters (longitudinal and marginal folding of petals) did not show marked differences among the proposed convarieties. However, differences among the convarieties in quantitative traits defining them (plant height, technical stem length and seed size) were considerable. Patterns of variation among the convarieties for other quantitative characters (petal width and seed oil content), as well as the frequencies of character expressions of four qualitative characters (petal colour, anther colour, ciliation of capsule septa and seed colour) were significantly associated with the four proposed convarieties, underlining the phenotypic and genotypic validity of this grouping. The patterns of geographic distribution of the convarieties and important characters showed that certain convarieties dominate in some areas of origin. The infraspecific classification and the presented characterization data increase the transparency of genetic diversity available in cultivated flax and in particular in the PGRC flax collection.

Ex situ collections of cultivated flax (Linum usitatissimum L.) and other species of the genus Linum L.

An overview of ex situ collections of cultivated flax (Linum usitatissimum L.) and 53 other species of the genus Linum L. is presented. The names of 33 genebanks in 23 countries preserving Linum germplasm and the species preserved at these places are provided. World genebanks engage in ex situ preservation of about 48,000 accessions of cultivated flax (Linum usitatissimum). However, possibly only 10,000 accessions of these are unique. Of the ca. 200 species of the genus Linum, other than L. usitatissimum, 53 are reported to be represented in the world’s ex situ collections with a total of about 900 accessions. Of these, 279 accessions alone refer to the wild progenitor of cultivated flax, Linum bienne Mill. However, L. bienne from the region of origin of cultivated flax is rarely represented in genebank collections. References to publications describing the techniques for ex situ conservation and the infraspecific variation of cultivated flax are made. It is suggested that taxonomic species delimitation within the genus Linum is in need of clarification and that the variation in Linum species for ornamental use or use in breeding is investigated.

Towards a better monitoring of seed ageing under ex situ seed conservation

Long-term conservation of 7.4 million ex situ seed accessions held in agricultural genebanks and botanic gardens worldwide is a challenging mission for human food security and ecosystem services. Recent advances in seed biology and genomics may have opened new opportunities for effective management of seed germplasm under long-term storage. Here, we review the current development of tools for assessing seed ageing and research advances in seed biology and genomics, with a focus on exploring their potential as better tools for monitoring of seed ageing. Seed ageing is found to be associated with the changes reflected in reactive oxygen species and mitochondria-triggered programmed cell deaths, expression of antioxidative genes and DNA and protein repair genes, chromosome telomere lengths, epigenetic regulation of related genes (microRNA and methylation) and altered organelle and nuclear genomes. Among these changes, the signals from mitochondrial and nuclear genomes may show the most promise for use in the development of tools to predict seed ageing. Non-destructive and non-invasive analyses of stored seeds through calorimetry or imaging techniques are also promising. It is clear that research into developing advanced tools for monitoring seed ageing to supplement traditional germination tests will be fruitful for effective conservation of ex situ seed germplasm.

Genomic regions underlying agronomic traits in linseed (Linum usitatissimum L.) as revealed by association mapping

The extreme climate of the Canadian Prairies poses a major challenge to improve yield. Although it is possible to breed for yield per se, focusing on yield-related traits could be advantageous because of their simpler genetic architecture. The Canadian flax core collection of 390 accessions was genotyped with 464 simple sequence repeat markers, and phenotypic data for nine agronomic traits including yield, bolls per area, 1,000 seed weight, seeds per boll, start of flowering, end of flowering, plant height, plant branching, and lodging collected from up to eight environments was used for association mapping. Based on a mixed model (principal component analysis (PCA) + kinship matrix (K)), 12 significant marker-trait associations for six agronomic traits were identified. Most of the associations were stable across environments as revealed by multivariate analyses. Statistical simulation for five markers associated with 1000 seed weight indicated that the favorable alleles have additive effects. None of the modern cultivars carried the five favorable alleles and the maximum number of four observed in any accessions was mostly in breeding lines. Our results confirmed the complex genetic architecture of yield-related traits and the inherent difficulties associated with their identification while illustrating the potential for improvement through marker-assisted selection.

Assembling a core collection from the flax world collection maintained by Plant Gene Resources of Canada

Between 1998 and 2008, numerous projects were conducted by the Canadian national genebank, Plant Gene Resources of Canada, for the regeneration, characterization and evaluation of the whole flax (Linum usitatissimum L.) germplasm collection. The whole collection comprised 3378 accessions and, according to the passport data, several of these were probably genetically very similar or even identical. Therefore, a subset of 381 accessions was selected that represented the diversity found in the whole collection. Sampling accessions from the whole collection was made using characterization and evaluation data and followed six different methods: (1) For seven qualitative characters, each unique combination of character expression was represented by three accessions; (2) for quantitative characters, a fixed number of accessions representing the lowest and highest observed values was included; (3) for stem fibre content, disease ratings, seed vigour and drought tolerance, a fixed number of accessions with desirable performance was included; (4) a subset of the 57 most distinct accessions based on RAPD markers was included; (5) a subset of 40 pure lines that were created based on extreme low and high values for 1000 seed weight, seed oil content and fatty acid profiles was included; (6) a subset of fibre flax cultivars of known relevance in European flax breeding and another subset of flax cultivars of known relevance for North American linseed breeding were included. The goal was to maximize the diversity available in a limited number of flax accessions by preserving the range of variation present in the whole collection, while improving evenness. The core collection was assembled in response to requests by flax breeders. This paper compares distribution parameters in the whole and core collections.

High-density marker profiling confirms ancestral genomes of Avena species and identifies D-genome chromosomes of hexaploid oat

Key messageGenome analysis of 27 oat species identifies ancestral groups, delineates the D genome, and identifies ancestral origin of 21 mapped chromosomes in hexaploid oat.AbstractWe investigated genomic relationships among 27 species of the genus Avena using high-density genetic markers revealed by genotyping-by-sequencing (GBS). Two methods of GBS analysis were used: one based on tag-level haplotypes that were previously mapped in cultivated hexaploid oat (A. sativa), and one intended to sample and enumerate tag-level haplotypes originating from all species under investigation. Qualitatively, both methods gave similar predictions regarding the clustering of species and shared ancestral genomes. Furthermore, results were consistent with previous phylogenies of the genus obtained with conventional approaches, supporting the robustness of whole genome GBS analysis. Evidence is presented to justify the final and definitive classification of the tetraploids A. insularis, A. maroccana (=A. magna), and A. murphyi as containing D-plus-C genomes, and not A-plus-C genomes, as is most often specified in past literature. Through electronic painting of the 21 chromosome representations in the hexaploid oat consensus map, we show how the relative frequency of matches between mapped hexaploid-derived haplotypes and AC (DC)-genome tetraploids vs. A- and C-genome diploids can accurately reveal the genome origin of all hexaploid chromosomes, including the approximate positions of inter-genome translocations. Evidence is provided that supports the continued classification of a diverged B genome in AB tetraploids, and it is confirmed that no extant A-genome diploids, including A. canariensis, are similar enough to the D genome of tetraploid and hexaploid oat to warrant consideration as a D-genome diploid.

Genetic and genomic resources of lentil: status, use and prospects

Extensive collections of lentil germplasm now exist in various genebanks around the world. This germplasm including wild Lens species has been used in plant introduction strategies and in efforts to widen the potential sources of increasing genetic diversity in the breeding programmes of lentil. Improved techniques are emerging to overcome hybridization barriers between species and as a result, interspecific hybrids have been successfully obtained between species. Several interspecific recombinant inbred line populations have been developed. Selected and backcrossed lentil lines are currently in advanced yield trial stages, and desirable traits such as yield, disease resistance and agronomic traits have been incorporated into cultivated lentil especially from Lens ervoides , generating a wider spectrum of variability. Secondly, further expansion of the overall pool of germplasm and examination of allelic variation at the nucleotide level will benefit lentil-breeding programmes by augmenting phenotype-based variation to further advance cultivar development. Genomic resources for lentil are limited now, but this situation is changing rapidly as the cost of genotyping has declined. As a result, two successive expressed sequence tags (EST) projects were undertaken under the NAPGEN EST project initiative ( http://www.nrc-cnrc.gc.ca/eng/programs/pbi/plant-products/napgen/.htm ) and an Agricultural Development Fund project initiative. We emphasize that creation of intraspecific and interspecific genetic populations, genetic maps, association maps, quantitative trait loci and marker-assisted selection technologies for implementation in the breeding programme will enhance deployment of genes responsible for traits of interest. The economical use of genomic technologies for use in germplasm resource management and genetic improvement is on the near horizon.