CRISPR/Cas9-Mediated Targeted Mutagenesis of CYP93E2 Modulates the Triterpene Saponin Biosynthesis in Medicago truncatula

In the Medicago genus, triterpene saponins are a group of bioactive compounds extensively studied for their different biological and pharmaceutical properties. In this work, the CRISPR/Cas9-based approach with two single-site guide RNAs was used in Medicago truncatula (barrel medic) to knock-out the CYP93E2 and CYP72A61 genes, which are responsible for the biosynthesis of soyasapogenol B, the most abundant soyasapogenol in Medicago spp. No transgenic plants carrying mutations in the target CYP72A61 gene were recovered while fifty-two putative CYP93E2 mutant plant lines were obtained following Agrobacterium tumefaciens-mediated transformation. Among these, the fifty-one sequenced plant lines give an editing efficiency of 84%. Sequencing revealed that these lines had various mutation patterns at the target sites. Four T0 mutant plant lines were further selected and examined for their sapogenin content and plant growth performance under greenhouse conditions. The results showed that all tested CYP93E2 knock-out mutants did not produce soyasapogenols in the leaves, stems and roots, and diverted the metabolic flux toward the production of valuable hemolytic sapogenins. No adverse influence was observed on the plant morphological features of CYP93E2 mutants under greenhouse conditions. In addition, differential expression of saponin pathway genes was observed in CYP93E2 mutants in comparison to the control. Our results provide new and interesting insights into the application of CRISPR/Cas9 for metabolic engineering of high-value compounds of plant origin and will be useful to investigate the physiological functions of saponins in planta

Antifeedant and insecticidal effects of alfalfa saponins in the management of the Japanese beetle Popillia japonica

Popillia japonica is a quarantine pest of priority interest for the EU, given its potentially important economic, social and environmental impacts. Alternative strategies to chemical methods are essential to limit its spread in newly infested areas with favourable climatic and environmental conditions. Saponins are biologically active molecules widely distributed in plants, displaying a well-known repellent activity combined with a mortality effect against insects. In this context, saponins were extracted from alfalfa Medicago sativa, where medicagenic and zanhic acid glycosides and Soyasaponin I were the most abundant compounds and used in the laboratory and semi-field experiments for treating leaves of susceptible host plants for P. japonica. Under laboratory conditions, a food deterrence effect and a significant mortality rate were observed using Corylus avellana leaves treated at increasing saponin concentrations, ranging from 1% to 5% w/v. Semi-field condition experiment supported the food deterrence effect, as a significant food preference was observed for untreated plants of Vitis vinifera compared to treated plants. The promising results obtained suggest that alfalfa saponins could represent a potential eco-friendly approach for Japanese beetle control

Identification of the Volatile Components of Galium verum L. and Cruciata leavipes Opiz from the Western Italian Alps

The chemical composition of the volatile fraction from Galium verum L. (leaves and flowers) and Cruciata laevipes Opiz (whole plant), Rubiaceae, was investigated. Samples from these two plant species were collected at full bloom in Val di Susa (Western Alps, Turin, Italy), distilled in a Clevenger-type apparatus, and analyzed by GC/FID and GC/MS. A total of more than 70 compounds were identified, making up 92%–98% of the total oil. Chemical investigation of their essential oils indicated a quite different composition between G. verum and C. laevipes, both in terms of the major constituents and the dominant chemical classes of the specialized metabolites. The most abundant compounds identified in the essential oils from G. verum were 2-methylbenzaldheyde (26.27%, corresponding to 11.59 μg/g of fresh plant material) in the leaves and germacrene D (27.70%; 61.63 μg/g) in the flowers. C. laevipes essential oils were instead characterized by two sesquiterpenes, namely β-caryophyllene (19.90%; 15.68 μg/g) and trans-muurola-4(15),5-diene (7.60%; 5.99 μg/g); two phenylpropanoids, benzyl alcohol (8.30%; 6.71 μg/g), and phenylacetaldehyde (7.74%; 6.26 μg/g); and the green-leaf alcohol cis-3-hexen-1-ol (9.69%; 7.84 μg/g). The ecological significance of the presence of such compounds is discussed

Work harassment

Il volume riunisce i risultati di un percorso di ricerca attuato a Verona, nell\u2019arco di due anni, in collaborazione tra Universit\ue0, CISL e CERIL (Centro Studi sulle Relazioni Industriali). Tale percorso ha inteso indagare, in alcune realt\ue0 produttive veronesi, fenomeni legati allo stress lavorativo e psicosociale, al benessere e al malessere organizzativi, al mobbing e alle molestie sul lavoro (work harassment). A partire dalla letteratura sull\u2019argomento e dalle definizioni che \ue8 possibile rintracciare in essa relativamente ai diversi fenomeni indagati, la ricerca ha inteso anche soffermarsi su possibili esperienze virtuose (le cosiddette buone pratiche) che le aziende hanno cercato di rendere operative al fine di aumentare il benessere al lavoro, e diminuire il malessere, di coloro che, in un\u2019organizzazione, spendono il proprio tempo, le proprie energie e i propri vissuti cognitivi ed emotivi

Il Joint Project: contesto, attori e motivazioni alla ricerca.

Favretto G., Albertini C.G., Sartori R., Bortolani E., Biazzi D., Il Joint Project: contesto, attori e motivazioni alla ricerca, 49-58

Introduzione. Il Joint Project, tra mondo della ricerca e mondo del lavoro.

Introduzione al volume di Favretto G., Albertini C.G., Sartori R., Bortolani E., Biazzi D. (2009). Work harassment. Benessere e malessere al lavoro tra stress, mobbing e pratiche organizzative, Franco Angeli, Milano, 191 pagin

The Antioxidant Activity of a Commercial and a Fractionated Phycocyanin on Human Skin Cells In Vitro

The protective effects for cells against chemical and UVA stress of a commercial phycocyanin (PC) for food use and a PC extracted from Arthrospira platensis (Spirulina) in phosphate buffer were assessed. The purity of the commercial PC, spectrophotometrically estimated as A620/A280 and confirmed by HPLC, was higher than that of the fractionated PC (2.0 vs. 1.5) but was twofold less concentrated. The oxygen radical antioxidant capacities (ORACs) of the commercial and fractionated PCs were 12,141 ± 1928 and 32,680 ± 3295 TE/100 g, respectively. The degradation of PCs upon exposure to UVA was spectrophotometrically estimated, and cytotoxicity was evaluated with the MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) test on human fibroblasts and keratinocytes. A lower level of reactive oxygen species (ROS) was recorded in the two cell lines incubated with the commercial PC after menadione treatment (p p p < 0.01) paradoxical prooxidant effects. Overall, the PC for food consumption demonstrated a high safety threshold and antioxidant ability to cells that, along with its coloring power, make it an excellent candidate for cosmetic formulations