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. Wellcharacterized 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.

Alkaloids represent the main antinutritional factor in lupins. The total content and qualitative composition of four major alkaloids in L. angustifolius L. were analyzed. The material included 329 accessions from the Polish collection divided into three classes of origin: wild lines, cultivars, and other manmade accessions. A very broad differentiation was found in terms of total alkaloid content-from 0.0005 to 2.8752 % of seed dry weight. In most cases, cultivars were characterized by a sharply decreased content of alkaloids, even below 0.01 % of seed dry weight. The average proportions of individual alkaloids were also very differentiated: lupanine-0.98 to 73.0 % of total alkaloid content (mean 46.4 %), 13-hydroxylupanine-15.6-71.1 % (mean 35.6 %), angustifoline-0-49.8 % (mean 15.5 %) and isolupanine-0-34.0 % (mean 2.5 %). The above mean values are probably typical for this species. In some accessions, lupanine and 13-hydroxylupanine accounted for 90-100 % of all major alkaloids. The average content of isolupanie (2.5 % of all alkaloids) allows it to be consider a major alkaloid of L. angustifolius, but quite frequently a ratio below 1 % or even its absence was stated. The three classes of origin were divided into three significantly different groups based on total alkaloid content as well as individual alkaloid content. Among wild lines, high alkaloid accessions were most numerous, but among cultivars it was low alkaloid accessions. The last class also contained numerous accessions with the lowest content of individual alkaloids. The influence of the content of individual alkaloids on total alkaloid content was also investigated in the wild lines and cultivars.

The total contents and qualitative compositions of alkaloids in seeds of 10 Old World lupin species (73 accessions) were surveyed using gas chromatography. The obtained results, combined with those for three lupin crops, Lupinus angustifolius, Lupinus albus, and Lupinus luteus, provide the most complete and up-to-date overview of alkaloid profiles of 13 lupin species originating from the Mediterranean Basin. The qualitative alkaloid compositions served as useful supplementary tools of species discrimination. On the basis of the most abundant major alkaloids, lupanine, lupinine, and multiflorine, the Old World lupin species were divided into four groups. Those containing lupanine (L. angustifolius, L. albus, and Lupinus mariae-josephi), containing lupinine (Lupinus luteus, Lupinus hispanicus, and Lupinus × hispanicoluteus), containing lupinine and multiflorine (Lupinus atlanticus, Lupinus palaestinus, Lupinus anatolicus, Lupinus digitatus, Lupinus pilosus, and Lupinus cosentinii), and containing multiflorine (Lupinus micranthus). Within a given group, certain species can be, in most cases, further distinguished by the presence of other major alkaloids. The discrimination of species based on the total alkaloid content was found to be less reliable because of the significant intra-species variations, as well as the influences of environmental factors on the seed alkaloid content.

Pea and lupin plants have been potentially rich sources of protein for human and animal diets. Currently, in Poland the main source of protein is soya cake, imported to Poland in amount of 2 mln tons per year. For substitution of soya with domestic protein plants breeding methods for higher and stable yield of grain legumes should be developed. About 12 generations are needed to develop a new cultivar and under central or north European climate conditions only one generation per year is feasible in the field. For that reason the development of new tools is needed to accelerate obtaining homozygosity in breeding of pea and lupins. Homozygosity can be attained by selfing successive generations or by hapoidization of hybrids using different in vitro techniques and doubling chromosomes in haploid plants (for review see, e.g., [1][2][3]). Pisum and Lupinus species are known to be recalcitrant to in vitro culture [4][5][6][7]. In spite of numerous studies focused on the production of doubled haploids (DH), no important results have been noted to date [8][9][10][11]. For shortening the breeding cycle, the single seed descent technique (SSD) in combination with in vitro culture of immature embryos may be an alternative to the DH system. Currently, SSD populations are frequently used as alternatives to DH populations in genetic and genomic studies (see, for example, [12,13]). The SSD technique connected with embryo in vitro culture allows the attainment of homozygous lines in a relatively short time. Such an approach applied to the development of winter barley SSD lines enabled the shortening of the generation cycle to ca. 4 months (including verbalization) [14]. Ochatt et al. [15] proposed the reduction of pea generation cycles based on in vitro culture of embryos and a greenhouse strategy, but no information was found on embryo culture for accelerating breeding process in lupins.The aim of the present studies was to establish in vitro conditions for the culture of pea, and narrow-leafed and yellow lupin embryos as the first step in research aimed at shortening generation cycles in the SSD technique used to accelerate the development of homozygous populations. Material and methodsMaterial for the studies consisted of two pea cultivars (Pisum sativum L.) -Cysterski and Akord, two narrow-leafed lupin cultivars (Lupinus angustifolius L.) -Kadryl and Sonet, and two yellow lupin cultivars (L. luteus L.) -Mister and Taper. Pea and lupin cultivar seeds were sown in the experimental field at Wiatrowo near Poznań. Pods from each pea and lupin cultivar were detached 21-28 days after flowering, when seeds were still-green but fully developed. Seeds taken out of pods were sterilized by successive dips into ethanol 70% (3 min) and AbstractThe aim of the studies was to establish in vitro conditions for the culture of pea and lupin embryos as the first step in the development of an in vitro assisted single seed descent technique for the attainment of homozygous populations. Materials for the study included of pea, and narrow-leafed a...

The narrow-leafed lupin, Lupinus angustifolius L., is a grain legume crop, cultivated both as a green manure and as a source of protein for animal feed and human food production. During its domestication process, numerous agronomic traits were improved, however, only two trait-related genes were identified hitherto, both by linkage mapping. Genome-wide association studies (GWAS), exploiting genomic sequencing, did not select any novel candidate gene. In the present study, an innovative method of 3 0-end reduced representation transcriptomic profiling, a massive analysis of cDNA ends, has been used for genotyping of 126 L. angustifolius lines surveyed by field phenotyping. Significant genotype × environment interactions were identified for all phenology and yield traits analysed. Principal component analysis of population structure evidenced European domestication bottlenecks, visualized by clustering of breeding materials and cultivars. GWAS provided contribution towards deciphering vernalization pathway in legumes, and, apart from highlighting known domestication loci (Ku/Julius and mol), designated novel candidate genes for L. angustifolius traits. Early phenology was associated with genes from vernalization, cold-responsiveness and phosphatidylinositol signalling pathways whereas high yield with genes controlling photosynthesis performance and abiotic stress (drought or heat) tolerance. PCR-based toolbox was developed and validated to enable tracking desired alleles in marker-assisted selection. Narrow-leafed lupin was genotyped with an innovative method of transcriptome profiling and phenotyped for phenology, growth and yield traits in field. Early phenology was found associated with genes from cold-response, vernalization and phosphatidylinositol signalling pathways, Piotr Plewi nski and Michał Książkiewicz contributed equally to the manuscript.