Cynara cardunculus L. var. scolymus L. Landrace "Carciofo Ortano" as a Source of Bioactive Compounds.

The preservation of agricultural biodiversity and socioeconomic development are relevant both to enhance domestic production and to support innovation. In the search for new biomolecules, we have focused on the "Carciofo Ortano" landrace, growth in the northern part of the Lazio region. Artichoke cultivation generates substantial by-products, including leaves, stems, and roots, which could serve as valuable sources of biomolecules and prebiotic dietary fiber. To valorize the leaf waste of the "Carciofo Ortano" landrace, a multidisciplinary approach was applied. Chemical analysis using HPLC-DAD identified mono-O- and di-O-caffeoylquinic acids and the sesquiterpene cynaropicrin in all artichoke leaf extracts. SPME-GC/MS analyses detected aliphatic alcohols in the fresh leaf samples. Antiproliferative and cytotoxic studies on cancer (SH-SY5Y, MCF-7, MDA) and normal (MCF-10A) human cell lines revealed that leaf extracts induced a selective dose and time-dependent biological effect. While showing slight activity against environmental bacterial strains, artichoke leaf extracts exhibited significant antifungal activity against the phytopathogenic fungus Alternaria alternata. Overall, the results highlight the potential of "Carciofo Ortano" cultivation by-products as a rich source of biomolecules with versatile applications in humans, animals, and the environment.

Morphological, Molecular, and Nutritional Characterisation of the Globe Artichoke Landrace "Carciofo Ortano".

The present study focused on the molecular, morphological, and nutritional characterisation of a globe artichoke landrace at risk of genetic erosion still cultivated in the municipality of Orte (Lazio Region, Central Italy) and therefore named "Carciofo Ortano". Molecular analysis based on SSR and ISSR markers was carried out on 73 genotypes selected at random from 20 smallholdings located in the Orte countryside and 17 accessions of landraces/clones belonging to the main varietal types cultivated in Italy. The results confirmed that "Carciofo Ortano" belongs to the "Romanesco" varietal typology and revealed the presence within the landrace of two distinct genetic populations named Orte 1 and Orte 2. Despite the high level of within-population genetic variation detected, the two populations were genetically differentiated from each other and from the landraces/clones of the main varietal types cultivated in Italy. Morphological and nutritional characterisation was performed on representative genotypes for each of the two populations of the "Carciofo Ortano" and the four landraces/clones included in the varietal platform of the PGI "CARCIOFO ROMANESCO DEL LAZIO" used as reference genotypes ("Campagnano", "Castellammare", "C3", and "Grato 1"). Principal component analysis showed that, of the 43 morphological descriptors considered, 12, including plant height, head shape index, head yield, and earliness, allowed a clear grouping of genotypes, distinguishing Orte 1 and Orte 2 populations from the reference genotypes. Regarding the nutritional composition of heads, particular attention should be devoted to the Orte 2 genotypes for their high dietary fibre, inulin, flavonoid, and phenol content, a feature that could be highly appreciated by the market.

Genetic Diversity and Population Structure of Common Bean (Phaseolus vulgaris L.) Landraces in the Lazio Region of Italy.

Common bean cultivation has historically been a typical component of rural economies in Italy, particularly in mountainous and hilly zones along the Apennine ridge of the central and southern regions, where the production is focused on local landraces cultivated by small-scale farmers using low-input production systems. Such landraces are at risk of genetic erosion because of the recent socioeconomic changes in rural communities. One hundred fourteen accessions belonging to 66 landraces still being grown in the Lazio region were characterized using a multidisciplinary approach. This approach included morphological (seed traits), biochemical (phaseolin and phytohemagglutinin patterns), and molecular (microsatellite loci) analyses to investigate their genetic variation, structure, and distinctiveness, which will be essential for the implementation of adequate ex situ and in situ conservation strategies. Another objective of this study was to determine the original gene pool (Andean and Mesoamerican) of the investigated landraces and to evaluate the cross-hybridization events between the two ancestral gene pools in the P. vulgaris germplasm in the Lazio region. Molecular analyses on 456 samples (four for each of the 114 accessions) revealed that the P. vulgaris germplasm in the Lazio region exhibited a high level of genetic diversity (He = 0.622) and that the Mesoamerican and Andean gene pools were clearly differentiated, with the Andean gene pool prevailing (77%) and 12% of landraces representing putative hybrids between the two gene pools. A model-based cluster analysis based on the molecular markers highlighted three main groups in agreement with the phaseolin patterns and growth habit of landraces. The combined utilisation of morphological, biochemical, and molecular data allowed for the differentiation of all landraces and the resolution of certain instances of homonymy and synonymy. Furthermore, although a high level of homozygosity was found across all landraces, 32 of the 66 examined (49%) exhibited genetic variability, indicating that the analysis based on a single or few plants per landrace, as usually carried out, may provide incomplete information.

Molecular markers dataset to assess the genetic diversity of oriental plane trees from historical sites in Lazio (central Italy).

The dataset presented in this article is related to the research paper titled "Dimensional and genetic characterization of the last oriental plane trees (Platanus orientalis L.) of historical sites in Lazio (central Italy)" (Ciaffi et al., 2022). Indeed, the molecular analyses reported in that article consisted in a comparison of Italian veteran plane trees with 12 certified accessions of P. orientalis, P. occidentalis and their hybrids P. acerifolia (4 individuals per species). First, LEAFY gene analyses allowed identifying 32 P. orientalis and two P. acerifolia in four sites of the province of Rome, confirming also that the two representative trees from the two gardens of the province of Viterbo belong to P. orientalis. Second, the use of Simple Sequence Repeat (SSR) and Inter Simple Sequence Repeat (ISSR) molecular markers provided useful information regarding the genetic relationships within and among all the historical sites. Owing to the use of SSR and ISSR molecular markers, a dataset of parameters related to the genetic diversity of the same plant material was obtained and presented in this article. For SSR markers, seven loci previously developed for P. occidentalis (Lang, 2010) and two specifically developed for P. orientalis (Rinaldi et al., 2019) were employed. For ISSR markers, DNA samples were amplified with eight primers before used for the determination of genetic stability of micro-propagated plantlets of P. acerifolia (Huang et al., 2009) and for the genetic characterization of plane trees within the formal gardens of Villa Lante of Bagnaia and Palazzo Farnese (Viterbo, Italy) (Ciaffi et al., 2018). To the best of our knowledge, this is the first report on the genetic diversity data for veteran oriental plane trees within heritage sites, which will offer helpful information for their management and conservation.

Monoterpene Synthase Genes and Monoterpene Profiles in Pinus nigra subsp. laricio.

In the present study, we carried out a quantitative analysis of the monoterpenes composition in different tissues of the non-model conifer Pinus nigra J.F. Arnold subsp. laricio Palib. ex Maire (P. laricio, in short). All the P. laricio tissues examined showed the presence of the same fourteen monoterpenes, among which the most abundant were ß-phellandrene, α-pinene, and ß-pinene, whose distribution was markedly tissue-specific. In parallel, from the same plant tissues, we isolated seven full-length cDNA transcripts coding for as many monoterpene synthases, each of which was found to be attributable to one of the seven phylogenetic groups in which the d1-clade of the canonical classification of plants' terpene synthases can be subdivided. The amino acid sequences deduced from the above cDNA transcripts allowed to predict their putative involvement in the biosynthesis of five of the monoterpenes identified. Transcripts profiling revealed a differential gene expression across the different tissues examined, and was found to be consistent with the corresponding metabolites profiles. The genomic organization of the seven isolated monoterpene synthase genes was also determined.

Diterpene Resin Acids and Olefins in Calabrian Pine (Pinus nigra subsp. laricio (Poiret) Maire) Oleoresin: GC-MS Profiling of Major Diterpenoids in Different Plant Organs, Molecular Identification and Expression Analysis of Diterpene Synthase Genes.

A quali-quantitative analysis of diterpenoid composition in tissues obtained from different organs of Pinus nigra subsp. laricio (Poiret) Maire (Calabrian pine) was carried out. Diterpene resin acids were the most abundant diterpenoids across all the examined tissues. The same nine diterpene resin acids were always found, with the abietane type prevailing on the pimarane type, although their quantitative distribution was found to be remarkably tissue-specific. The scrutiny of the available literature revealed species specificity as well. A phylogeny-based approach allowed us to isolate four cDNAs coding for diterpene synthases in Calabrian pine, each of which belonging to one of the four groups into which the d3 clade of the plants' terpene synthases family can be divided. The deduced amino acid sequences allowed predicting that both monofunctional and bifunctional diterpene synthases are involved in the biosynthesis of diterpene resin acids in Calabrian pine. Transcript profiling revealed differential expression across the different tissues and was found to be consistent with the corresponding diterpenoid profiles. The isolation of the complete genomic sequences and the determination of their exon/intron structures allowed us to place the diterpene synthase genes from Calabrian pine on the background of current ideas on the functional evolution of diterpene synthases in Gymnosperms.

Profiling Volatile Terpenoids from Calabrian Pine Stands Infested by the Pine Processionary Moth.

Terpenoids make up the biggest and most diversified class of chemical substances discovered in plants, encompassing over 40,000 individual compounds. In conifers, the production of terpenoids, either as oleoresin or emitted as volatile compounds, play an important role in the physical and chemical defence responses against pathogens and herbivores. In the present work, we examined, for the first time to the best of our knowledge, the terpenic defensive relations of Calabrian pine (Pinus nigra subsp. laricio (Poiret) Maire), facing the attack of the pine processionary moth (Thaumetopoea pityocampa (Denis and Schiffermüller, 1775)), brought about in the open on adult plant individuals growing at two distinct forest sites. Among the volatile terpenoids emitted from pine needles, bornyl acetate [(4,7,7-trimethyl-3-bicyclo[2.2.1]heptanyl) acetate] was the most frequently and selectively associated with the infestation, increasing during the period of most intense trophic activity of the caterpillars (defoliation), and decreasing thereafter. Although further work is needed to clarify whether the observed response reflects defence reactions and/or they are involved in communication among the infested plants and their biotic environment, the present results boost the currently growing interest in the isolation and characterization of plant secondary metabolites that can be used to control pests, pathogens, and weeds.

On the Evolution and Functional Diversity of Terpene Synthases in the Pinus Species: A Review.

In the biosynthesis of terpenoids, the ample catalytic versatility of terpene synthases (TPS) allows the formation of thousands of different molecules. A steadily increasing number of sequenced plant genomes invariably show that the TPS gene family is medium to large in size, comprising from 30 to 100 functional members. In conifers, TPSs belonging to the gymnosperm-specific TPS-d subfamily produce a complex mixture of mono-, sesqui-, and diterpenoid specialized metabolites, which are found in volatile emissions and oleoresin secretions. Such substances are involved in the defence against pathogens and herbivores and can help to protect against abiotic stress. Oleoresin terpenoids can be also profitably used in a number of different fields, from traditional and modern medicine to fine chemicals, fragrances, and flavours, and, in the last years, in biorefinery too. In the present work, after summarizing the current views on the biosynthesis and biological functions of terpenoids, recent advances on the evolution and functional diversification of plant TPSs are reviewed, with a focus on gymnosperms. In such context, an extensive characterization and phylogeny of all the known TPSs from different Pinus species is reported, which, for such genus, can be seen as the first effort to explore the evolutionary history of the large family of TPS genes involved in specialized metabolism. Finally, an approach is described in which the phylogeny of TPSs in Pinus spp. has been exploited to isolate for the first time mono-TPS sequences from Pinus nigra subsp. laricio, an ecologically important endemic pine in the Mediterranean area.

Transcriptional regulation of stilbene synthases in grapevine germplasm differentially susceptible to downy mildew.

BACKGROUND: To limit the impact of the downy mildew disease of grapevine and reduce the need to recur to chemical treatments, an effective strategy might be recovering adaptive resistance traits in both cultivated and wild V. vinifera germplasm. Considering that stilbenes represent the most important class of phytoalexins in the Vitaceae, the constitutive expression and transcriptional activation of all the functional members of the stilbene synthase gene family were analysed in a group of nine grapevine genotypes following artificial infection with the oomycete Plasmopara viticola, the causal agent of the disease. In addition, in the same genotypes we analyzed the expression of genes encoding for two transcription factors involved in the transcriptional regulation of the stilbene synthase genes, namely VvMYB14 and VvMYB15, and of genes encoding for chalcone synthases. RESULTS: Downy mildew incidence and severity ranged from nihil to high in the grapevine genotypes considered, being low to moderate in a subgroup of V. vinifera genotypes. The constitutive expression of the stilbene synthase genes as well as the extent of their transcriptional activation following P. viticola inoculation appeared to be inversely related to the proneness to develop disease symptoms upon infection. In a specular manner, following P. viticola inoculation all the chalcone synthase genes were up-regulated in the susceptible grapevine genotypes and down-regulated in the resistant ones. The infection brought by P. viticola appeared to elicit a co-ordinated and sequential transcriptional activation of distinct stilbene synthase genes subsets, each of which may be regulated by a distinct and specific MYB transcription factor. CONCLUSIONS: The present results suggest that the induction of stilbene biosynthesis may contribute to the basal immunity against the downy mildew of grapevine, thus representing an adaptive resistance trait to recover, in both cultivated and wild V. vinifera germplasm. During the early stages of P. viticola infection, an antagonistic interaction between flavonol and stilbene biosynthesis might occur, whose outcome might determine the subsequent extent of disease symptoms. Further studies are needed to decipher the possible regulatory mechanisms involved in the antagonistic crosstalk between these two metabolic pathways in resistant and susceptible genotypes in response to P. viticola.

Data set useful for the micropropagation and the assessment of post-vitro genetic fidelity of veteran trees of P. orientalis L.

The data presented in this article are related to the research article titled "Conservation of veteran trees within historical gardens (COVE): a case study applied to Platanus orientalis L. in central Italy" (Ciaffi et al., 2018) [1]. This article reports data on the composition of the substrates used in the different steps of Platanus orientalis micropropagation: establishment of in vitro culture, multiplication, elongation and rooting. Moreover, molecular data were used to assess the genetic fidelity of the micropropagated plants respect mother plants after three year of in vitro cultivation. Fifteen ISSR markers, used in "Determination of genetic stability of long-term micropropagated plantlets of Platanus acerifolia using ISSR markers" (Huang et al., 2009) [2] on P. acerifolia and in "Variant identification in Platanus occidentalis L. using SNP and ISSR markers" (Lee et al., 2012) [3] on P. occidentalis, were successfully employed in the present study on P. orientalis. The plant material was collected from the Renaissance garden of Villa Lante in Viterbo, Italy. It is envisioned that these data set will provide useful information for the conservation of veteran oriental plane trees of historical gardens.