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Milk thistle, Silybum marianum (L.) Gaertn. (Asteraceae), is an economically important medicinal plant utilized for silymarin production. Moreover, the species has been positively evaluated for vegetable oil and biomass production. Despite these positive characteristics, milk thistle is still marked by traits that are typical of undomesticated species (most importantly natural fruit dispersal at maturity) and requires further genetic improvement for its complete exploitation. This manuscript summarizes all the information collected through time about a collection of nine milk thistle wild and mutant lines and it discusses the possible further utilization of these genotypes. The accessions are characterized by interesting traits related to: fruit silymarin composition (S. marianum chemotype A and B), fruit fatty acid composition (high oleic and high stearic acid lines), fruit condensed tannins content, vegetative biomass composition (modification of xylans or lignin content), vegetative biomass structure (dwarf and tall lines), modifications of leaf variegation (hypervareigated line) and different types of fruit shatter resistance at maturity. All the lines underwent subsequent generations of selfing and are stable for all the described traits. The accessions will be made available at the Genebank of the Leibniz Institute of Plant Genetics and Crop Plant Research (IPK, Gatersleben) and may prove to be a useful genetic material for the improvement of qualitative fruit traits (silymarin quality, fatty acid composition) and for the further development of shatter-resistant S. marianum genotypes towards the complete domestication of this promising species.

Silybum marianum (L.) Gaertn. (milk thistle) is plant species that has been utilized principally for medicinal purposes for more than 2000 years. Recently it was proposed for biomass production in marginal environments, but vegetative biomass compositional analyses had not been available so far. The study of plant morphology and biomass composition was conducted on three different S. marianum accessions grown under open field conditions. The results indicate that plant morphological traits show major differences between accessions: this suggests that the available natural variability can be further utilized in order to develop improved S. marianum cultivars. Biomass compositional analysis shows that extractives, ash, lignin and cellulose content are comparable to other herbaceous bioenergy crops and that these traits display only limited variability in the studied accessions. Hemicellulose fraction is composed only by xylans and its content appears averagely lower in comparison to other herbaceous biomasses. Interestingly, in S. marianum biomass total nitrogen content is lower if compared to other herbaceous species. The possible involvement of this specific biomass trait in S. marianum nitrogen utilization efficiency has to be further investigated.

Milk thistle [Silybum marianum (L.) Gaertn.] is used as a medicinal plant for the production of silymarin. Other than silymarin, the fruit also contains valuable compounds such as oil and protein, and the lignocellulosic biomass of this species has been positively evaluated for biogas, syngas, and energy production. Despite these positive characteristics, the plant is still marked by traits that are typical of undomesticated species. Specifically, fruit shattering at maturity severely hampers its possible utilization as a crop. Therefore, for the first time, an ethyl methanesulfonate mutagenesis program was implemented to identify milk thistle lines with reduced fruit shattering, and its effectiveness was evaluated through genome-wide analysis. The candidate nonshattering lines were multiplied up to M₆ for the validation of the favorable traits. The lignocellulosic biomass of the selected shatter resistant lines was analyzed according to compositional analysis. The natural milk thistle fruit dispersal mechanism was shown to be modulated by changing air relative humidity. Three vigorous, stable, shatter-resistant lines were identified. This is the first report that describes milk thistle nonshattering lines. At this stage in the three lines, the morphological description and the compositional analysis of the lignocellulosic biomass indicated that shatter resistance was concomitant to changes in vegetative biomass structure and composition. This observation is compatible with the described natural fruit dispersal mechanism, and it is of interest for the design of new milk thistle crop ideotypes for the complete domestication of the species.

Transgenerational epigenetics inheritance refers to the possibility to inherit epigenetic-based information acquired from previous generations. The design of experiments that can measure this phenomenon presents complexities. These are related mainly to difficulties in the identification of epigenetic variation components that are independent from genetic variation, and to difficulties related to the time needed to expose genetically stable plants to different environments for several generations. In this study we propose an experimental approach that takes advantage of seed material routinely produced by seedbanks in order to circumvent the abovementioned issues. By taking advantage of this freely available seed material, it is possible to evaluate epigenetic differences induced by the environment experienced by previous generations and to evaluate both the extent and the impact of epigenetic variation in crops. Seeds of two barley (Hordeum vulgare L.) cultivars, which were previously multiplied in six different seedbanks located worldwide, were grown in a common environment and the plants were evaluated at a genetic, epigenetic, and phenotypic level. Our results indicate that barley accessions multiplied in different environments displayed a comparable level of epigenetic variation, which was, however, greater than the genetic variation. Cluster analysis of methylation differences, likewise DNA differences, split the two cultivars in two distinct groups evidencing a clear link between genetic and epigenetic variation. Nevertheless, the analysis of phenotypic traits and of the CCGG sites that are variable in both cultivars suggests that there might be a possible link between previous cultivation environment and induced epigenetic changes at specific DNA regions.

Silybum marianum (L.) Gaertn (Asteraceae) is a valuable medicinal plant utilized for silymarin production. However, only fragmentary and contradictory information about silymarin localization within S. marianum fruit are available. In this work, a twofold research approach was adopted in order to investigate the distribution and quantification of silymarin and of other phenolic compounds within the different fruit regions (pericarp, seed integument, cotyledon). Two S. marianum wild accessions with contrasting silymarin chemotype (A and B) and a mutant line (C) with an altered fruit colour were analysed. Fruits of Cynara cardunculus were studied as a reference. Firstly, the fruit morpho-anatomy was reviewed by means of light microscopy digital imaging and, secondly, a comprehensive histolocalization of the different classes of polyphenols within the fruit was carried out. The experimental evidences confirmed that silymarin, and its precursor taxifolin, are only accumulated in the seed integuments. The dark colour of fully-ripened fruits is due to the accumulation of condensed tannins in the pericarp subepidermal cell layer. On the contrary, the studied mutant line shows reduced condensed tannin content that probably result from impairment at the level of flavonoid biosynthetic pathway. Condensed tannins content is comparatively low in S. marianum fruits and very low in the identified mutant line. This could represent an advantage for the possible employment of S. marianum fruits and of silymarin extraction by-products in the feed and food sector.

We have developed a new version of the cTBP (combinatorial tubulin-based polymorphism) method, a previously described approach based on intron-length polymorphism (ILP), to rapidly characterize the β-tubulin gene family of Camelina sativa (L.) Crantz, a plant species of importance for oil production but still largely unexplored at genomic level. The method, named h-TBP, allows the rapid cloning of the β-tubulin genomic sequences that encompass the two introns, invariantly present at fixed positions within the coding region of the vast majority of the plant species. The β-tubulin sequences cloned by h-TBP also comprise part of exon1 and exon3 and the whole sequence of exon2. The h-TBP method has then been used to isolate, clone and characterize the β-tubulin gene family of C. sativa, composed of at least 20 different β-tubulin isotypes, named CsTUB1 through CsTUB20. The relatively high number of β-tubulin genes has been further substantiated by Southern-blot analysis. Comparison of the β-tubulin exon sequences of C. sativa with those of Arabidopsis thaliana, the closest relative among crucifers, defines distinct groups of putative orthologous genes, identified by a UPMGA cluster analysis. Analysis of the C. sativa β-tubulin intron sequences reveals some molecular features that can provide the first hints for the understanding of intron plasticity and evolution. From a more immediate perspective, these data provide the first substantial contribution to the characterization of the largely unexplored genome of C. sativa, and the tools for assisting programmes of breeding and selection of the most productive plants.

Oleander (Nerium oleander L.) is an evergreen shrub of great ornamental interest which, in recent times, has been increasingly used as a flowering pot plant. Plants grown in pots undergo more frequent water stress conditions than those grown in the soil, due to the limited volume of substrate available for the roots. Oleander is a species adaptable to dry conditions and able to survive long periods of drought. It is well known that under water stress conditions all plants reduce photosynthetic activity, resulting in reduced plant growth. In case of severe water stress, leaves undergo strong wilting and senescence resulting in the loss of ornamental value. In the present work, a study was conducted to evaluate the ecophysiological response to water stress in four oleander cultivars previously recognised (on the basis of traits such as size, habit, earliness, abundance and duration of flowering, aptitude for cutting propagation and rapidity of growth) as suitable for pot plant production. Our data confirm the high drought tolerance of oleander. In the studied cultivars, plants submitted to water stress showed only minor variations in leaf gas exchange parameters [transpiration (E), stomatal conductance (gs) and CO2 net assimilation (A)] for at least 10 days without a change in leaf water content [assessed as relative water content (RWC)] for 22 days from the beginning of the stress treatment. During this period, non-irrigated plants maintained the same water status as control plants and were visually undistinguishable from them. Moreover, plants survived without water supply for one month. Following the supply of water again, they were able to restore RWC, gas exchange parameters and instantaneous water use efficiency [A/E ratio (WUEinst)] to the values of control plants. Furthermore, if at the end of the stress period plants appeared withered and were pale green in colour, they regained their normal appearance after they were irrigated again. Although the four studied cultivars showed some minor differences in leaf gas exchange parameters and in the manner in which the latter parameters changed after irrigation was stopped, the response to water stress was essentially the same. Therefore, as far as drought tolerance is concerned, all these cultivars have a good aptitude for use as flowering pot plants.

Milk thistle [Silybum marianum (L.) Gaertn.] is an officinal crop species native to the Mediterranean. The species is gaining increasing interest as a multipurpose crop, but lack of information exists about the natural phenotypic variability available in the species for the traits of interest. In the present work, the fruit morphological and chemical traits of a collection of 26 milk thistle accessions have been analyzed. The results show that the most variable traits are related to single flavonolignan content. When the most abundant fatty acids present in the fruit are considered, oleic acid appears to have the higher variability between accessions. Correlations between variables highlight strong relationships between the different flavonolignans and a strong positive correlation between isosilybin A and the total silymarin content, suggesting a central role of this minor flavonolignan in silymarin biosynthetic pathway. The clustering procedure displays considerable diversity between the accessions that originate from localities where milk thistle is described as a native species.

BACKGROUND: Camelina sativa (CS) is an oilseed crop used for biofuel production. By-products from oil extraction are high in protein and can be used in ruminant rations; more information about their nutritive value is required also considering the antinutrional factor content of the by-products. The aim of this study was to evaluate the nutritive value of CSmeal genotypes in comparison with canola. RESULTS: TenCSgenotypesandonecanola cultivar were evaluated.Meals were obtained from seeds after solventoil extraction. CS average crude protein (CP) content (g kg-1 drymatter) was 457. Numerical differences in lysine and sulfur amino acid content were observed among CS genotypes. Glucosinolate (mmol kg-1) content was higher for CS (23.1) than canola (7.2). Sinapine content (g kg-1) was lower for CS (2.79) than for canola (4.32). Differences were observed among CS genotypes for rumen undegraded protein (RUP). Average RUP (g kg-1 CP) was 316 for CS and 275 for canola. CONCLUSIONS: CS meal has potential for use in ruminant rations as a high-quality protein source. In vivo studies are needed to compare CS with other protein sources used in cattle rations. Implementation of breeding programs for improved meal quality is recommend.