Wojciech Święcicki

New evidence of ancestral polyploidy in the Genistoid legume Lupinus angustifolius L. (narrow-leafed lupin)

Key messageThis is the first clear evidence of duplication and/or triplication of large chromosomal regions in a genome of a Genistoid legume, the most basal clade of Papilionoid legumes.AbstractLupinus angustifolius L. (narrow-leafed lupin) is the most widely cultivated species of Genistoid legume, grown for its high-protein grain. As a member of this most basal clade of Papilionoid legumes, L. angustifolius serves as a useful model for exploring legume genome evolution. Here, we report an improved reference genetic map of L. angustifolius comprising 1207 loci, including 299 newly developed Diversity Arrays Technology markers and 54 new gene-based PCR markers. A comparison between the L. angustifolius and Medicago truncatula genomes was performed using 394 sequence-tagged site markers acting as bridging points between the two genomes. The improved L. angustifolius genetic map, the updated M. truncatula genome assembly and the increased number of bridging points between the genomes together substantially enhanced the resolution of synteny and chromosomal colinearity between these genomes compared to previous reports. While a high degree of syntenic fragmentation was observed that was consistent with the large evolutionary distance between the L. angustifolius and M. truncatula genomes, there were striking examples of conserved colinearity of loci between these genomes. Compelling evidence was found of large-scale duplication and/or triplication in the L. angustifolius genome, consistent with one or more ancestral polyploidy events.

Genetic diversity in European Pisum germplasm collections

The distinctness of, and overlap between, pea genotypes held in several Pisum germplasm collections has been used to determine their relatedness and to test previous ideas about the genetic diversity of Pisum. Our characterisation of genetic diversity among 4,538 Pisum accessions held in 7 European Genebanks has identified sources of novel genetic variation, and both reinforces and refines previous interpretations of the overall structure of genetic diversity in Pisum. Molecular marker analysis was based upon the presence/absence of polymorphism of retrotransposon insertions scored by a high-throughput microarray and SSAP approaches. We conclude that the diversity of Pisum constitutes a broad continuum, with graded differentiation into sub-populations which display various degrees of distinctness. The most distinct genetic groups correspond to the named taxa while the cultivars and landraces of Pisum sativum can be divided into two broad types, one of which is strongly enriched for modern cultivars. The addition of germplasm sets from six European Genebanks, chosen to represent high diversity, to a single collection previously studied with these markers resulted in modest additions to the overall diversity observed, suggesting that the great majority of the total genetic diversity collected for the Pisum genus has now been described. Two interesting sources of novel genetic variation have been identified. Finally, we have proposed reference sets of core accessions with a range of sample sizes to represent Pisum diversity for the future study and exploitation by researchers and breeders.

An analysis of isozymic loci polymorphism in the core collection of the Polish Pisum genebank

The investigated material consisted of the core collection of the Polish Pisum gene bank at Wiatrowo and represented a described, monohybrid variation (type lines for characters/genes and their initial lines, tester lines with markers and lines from wild taxa, 266 accessions in total). A polymorphism in 18 isozymic loci was analyzed (Lap-1, Px-1, Aat-p, Aat-m, Aat-c, Pgm-c, Pgm-p, Gpi-c, Idh, Acp-1, Skdh, Est-1, Est-2, Est-3, Aldo, 6Pgd-p, 6Pgd-c, Dia-1). The practical outcome of the analysis was a selection of lines with a high number of fast or slow isozymic alleles, useful as tester lines with markers for chromosome mapping. A polymorphism was stated in all investigated loci with 2 to 4 alleles ranging from very fast, fast, slow to very slow. Fast and slow alleles occurred in all loci. In 11 loci the incidence of fast alleles was higher than slow ones. Conversly, slow alleles occurred more frequently than fast ones in 5 loci. Additionally, there was a low incidence of a very fast allele in 7 loci and of a very slow allele in 3 loci. A slow allele was rare in 2 loci. Primitive lines and land races were a source of these rare alleles. It is suggested to consider a frequent allele of a given locus found in a group of wild lines as a so-called wild type of Pisum.

Quantitative and qualitative analysis of alkaloids composition in the seeds of a white lupin (Lupinus albus L.) collection

White lupin (Lupinus albus L.) has unexploited potential as a crop plant due to its high seed yield as well as protein and oil content in seeds. Well-characterized collections of gene resources are very important for breeding as a source of genetic variation. This paper presents the results of analyses for total content and qualitative composition of alkaloids in seeds of 367 L. albus accessions from the Polish Genebank. Accessions were divided into four classes of origin: wild collected material, land races, breeding lines, and cultivars. Apart from the expected broad variation as well as strong differentiation in the alkaloid content, a clear influence of domestication was observed. This was shown as an apparent decrease in the alkaloid content in breeding lines and cultivars classes. The total alkaloid content varies from 0.02 to 12.73% of the seed dry weight. Six major alkaloids (abundance >1%) were revealed: lupanine (28.22–94.49%, mean 76.06% in total content), 13-hydroxylupanine (0.10–32.78%, mean 8.23%), multiflorine (0.00–21.67%, mean 5.52%), albine (0.00–18.55%, mean 4.48%), angustifoline (0.24–12.14%, mean 2.07%), 11,12-seco-12,13-didehydromultiflorine (0.00–12.28%, mean 1.74%). Owing to its abundance, lupanine was found to be the most closely correlated to the total alkaloid content.

Raffinose family oligosaccharides in seeds of Pisum wild taxa, type lines for seed genes, domesticated and advanced breeding materials

The content of raffinose family oligosaccharides (RFOs) in pea seeds constrains their usage in feeding humans and animals. In our research, the content of soluble carbohydrates—particularly α-d-galactosides of sucrose (RFOs)—was analyzed. The materials were as follows: 248 accessions from the Polish Pisum Genebank including representatives of taxa (from species to convarietas), type lines for genes controlling seed characters, and breeding materials and cultivars. Accessions were divided into homogeneous groups considering content of total soluble carbohydrates, total RFOs and individual RFOs: raffinose, stachyose and verbascose. The highest content of total soluble carbohydrates and total RFOs were stated for accessions with wrinkled seeds (r and rb genes) and the lowest content for seeds of the wild species P. fulvum Sibth. et Sm. Accessions valuable for breeding (for further decreasing of anti-nutritional compounds) were found among domesticated taxa (P. sativum subsp. sativum convar. vulgare Alef. and speciosum (Dierb.) Alef.), breeding lines, and some wild taxa. Accessions with decreased content of a total RFOs and verbascose are particularly valuable. It was found that the content of total RFOs was the most highly, frequently, and positively correlated with a stachyose and verbascose content. However, in P. fulvum seeds with the lowest content of RFOs and verbascose, a high correlation between the content of total RFOs and stachyose was revealed. Contents of all oligosaccharides were substantially lower in lines with dominant alleles of main pea seed genes (R, A, and I). It can be assumed that wild, primitive peas were characterized by not-all-to-high (rather not high) content of oligosaccharides; then recessive mutations in mentioned genes resulted in an increased content of RFOs. It seems to be an interesting observation from an evolutionary point of view.

Application of the High Resolution Melting analysis for genetic mapping of Sequence Tagged Site markers in narrow-leafed lupin (Lupinus angustifolius L.)

Sequence tagged site (STS) markers are valuable tools for genetic and physical mapping that can be successfully used in comparative analyses among related species. Current challenges for molecular markers genotyping in plants include the lack of fast, sensitive and inexpensive methods suitable for sequence variant detection. In contrast, high resolution melting (HRM) is a simple and high-throughput assay, which has been widely applied in sequence polymorphism identification as well as in the studies of genetic variability and genotyping. The present study is the first attempt to use the HRM analysis to genotype STS markers in narrow-leafed lupin (Lupinus angustifolius L.). The sensitivity and utility of this method was confirmed by the sequence polymorphism detection based on melting curve profiles in the parental genotypes and progeny of the narrow-leafed lupin mapping population. Application of different approaches, including amplicon size and a simulated heterozygote analysis, has allowed for successful genetic mapping of 16 new STS markers in the narrow-leafed lupin genome.

Professor Ignacy Wiatroszak (1924–2005)

who was the former head of the Institute of Plant Genetics in Poznañ and Deputy Editor-in-Chief of the Journal of Applied Genetics, died on 1 May 2005. He was born on 20 December 1924 in Dzier¿anów Wielkopolski in central Poland. He graduated from the Agricultural University in Poznañ in 1957, when he received a master’s degree for his thesis “Effect of Drought in Various Developmental Stages on the Oil Content of Some Oil Crops”. In 1959–1962 he was a researcher at the Experimental Laboratory of the Provincial Department of Animal Insemination in Poznañ. In 1963 he started to work at the Agricultural University in Poznañ. In 1965 he defended his PhD thesis “Variability of Blood Cell Antigens in Cattle Bred in Wielkopolska” and earned a doctoral degree in agricultural sciences (specialization: immunogenetics). Six years later, he finished his post-doctoral thesis “Immunogenetic Characteristics of Pigs Bred in Poland”. In 1972 he moved from the Agricultural University to the Department of Plant Genetics (now called the Institute of Plant Genetics), Polish Academy of Sciences, Poznañ. He became the head of the Department when Prof. Stefan Barbacki retired in 1973. He held this post till 1985 and again in 1990–1993. In 1976 he was awarded the scientific title of professor. The scientific research of Prof. Ignacy Wiatroszak covered various aspects of plant and animal genetics, with particular reference to the needs of agriculture. At the first stage of his investigations (1966–1972), his experiments focused on the use of blood group genes as well as proteins and enzymes in blood serum as markers of physiological and biochemical processes in animal organisms. These were then pioneering works, which arouse wide interest in Poland and other countries. Results of the investigations were presented at international congresses in Paris (1966), Warsaw (1968), Budapest (1970), and Vienna (1972). The second stage of his scientific research was concerned with theoretical aspects of genetics and breeding of crops. Results of his research were presented in several dozen publications. Special attention should be given to Prof. Wiatroszak’s achievements in the field of organization of scientific research. He transformed the Department into the Institute of Plant Genetics. He also coordinated research within the framework of central governmental programmes: “Breeding of High-Yielding Cultivars of Leguminous Plants” (1976–1980), “Genetic Shaping of Plant Productivity” (1981–1985), and “Genetic Improvement of Cultivated Plants” (1986–1990). Besides, in 1980–1994, he was Deputy Editor-inChief of the Journal of Applied Genetics, which is published by the Institute. Because of the great demand for genetic research on cultivated plants, Prof. Wiatroszak inspired the realization and coordination of several important research programmes, such as the longterm programme “Genetic Improvement of Cultivated Plants with the Use of Biotechnology and Genetic Engineering”. Achievements of that programme include the generation of many new genotypes that were used as donors for breeding of new cultivars, as well as the elaboration of new methods of early and effective selection of crop plants. In 1978–1983, Prof. Ignacy Wiatroszak performed the functions of the head of the Scientific Board of the Programme for Development of Plant Breeding and Seed Production in Poland in cooperation with the UNDP and the Polish Ministry of Agriculture. He was also the head of Scientific Boards at the Potato Institute in Bonin (1974–1985), the Institute of Plant Breeding and Acclimatization at Radzików (1985–1991), J Appl Genet 47(1), 2006, pp. 1–2

Preliminary Estimation of Variation of Alkaloids Content in White Lupin (Lupinus albus L.) Collection

Grain legume crops are crucial elements of global agriculture and nutrition, both as food and feed, since they are major sources of plant protein. Of the many species of genus Lupinus, L. angustifolius, L. luteus and L. albus (characterized by high protein content) are particularly important from the agricultural point of view. Among them, white lupin (L. albus) has unexploited potential in Poland as a crop plant due to its high seed yield as well as protein and oil content in the seeds. In white lupin breeding, the reduction of alkaloid content in seeds is important. The paper presents an assessment of the variability in total alkaloid content and qualitative composition of alkaloids in seeds of white lupin. Plant material constituted 150 accessions from domestic collections, divided into five classes: wild collected material, landraces, induced mutants, breeding lines, and cultivars. Total alkaloid content was analysed along with qualitative composition expressed by four major alkaloids, and broad variation was found. The total alkaloid content varied from 0.008 to 11.1% of seed dry weight. An apparent decrease in alkaloid content was shown in breeding lines and cultivars, indicating a clear influence of domestication. The four major alkaloids (abundance >1%) were lupanine (mean 71.3%), 13–hydroxylupanine (10.46%), multiflorine (6.8%) and angustifoline (3.8%). The lower alkaloid content of breeding lines and cultivars, as compared to wild lines and landraces, was associated with higher relative contents of 13–hydroxylupanine and angustifoline and lower relative contents of lupanine and multiflorine.

Effects of Temperature on Growth During in vitro Embryo Culture of Field Bean (Vicia faba var. minor L.)

Attainment of homozygous lines in grain legumes is multi-year procedure. It may be shortened by applying the single seed descent technique under greenhouse conditions. Additionally, this technique can be combined with in vitro culture of embryos dissected from immature seeds. The aim of this study was to establish in vitro conditions for the culture of field bean embryos as the first step in the development of homozygous lines. Four temperature regimes were applied and their influence on embryo development was evaluated. Under ex vitro conditions plant survival was observed. Our results indicate a good potential of embryo culture to shorten the time required for obtaining homozygous lines of field bean.

The fa2 gene and molecular markers mapping in the gp segment of the Pisum linkage group V

Review studies on the world Pisum genetic resources have shown that stem fasciation is controlled by three loci, i.e., fa1 (LGIV; Wt 10006 - type line of the Polish Gene Bank), fa2 (LGV, the line Wt 12185), and fas (LGIII, the line Shtambovii). Outstanding advantages of this character (e.g., pods gathered in upper part of a stem) resulted in breeding some cultivars. Preliminary investigations suggested linkages of the newly described fa2 gene within the gp–U segment. Based on the further linkage test crosses, it was stated that the fa2 is localized between the gp and Pis_Gen_9_3_1 markers (in the LGV). Additionally, four molecular markers (AD175, AB146, AC58, and AD280) and the morphological marker lk were also localized in this segment. Moreover, rms5, lum3, and cri were found to map on the other side of gp with tight linkage observed between lum3 and cri.