Manipulating the Biosynthesis of Bioactive Compound Alkaloids for Next-Generation Metabolic Engineering in Opium Poppy Using CRISPR-Cas 9 Genome Editing Technology

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated9 (Cas9) endonuclease system is a powerful RNA-guided genome editing tool. CRISPR/Cas9 has been well studied in model plant species for targeted genome editing. However, few studies have been reported on plant species without whole genome sequence information. Currently, no study has been performed to manipulate metabolic pathways using CRISPR/Cas9. In this study, the type II CRISPR/SpCas9 system was used to knock out, via nonhomologous end-joining genome repair, the 4′OMT2 in opium poppy (Papaver somniferum L.), a gene which regulates the biosythesis of benzylisoquinoline alkaloids (BIAs). For sgRNA transcription, viral-based TRV and synthetic binary plasmids were designed and delivered into plant cells with a Cas9 encoding-synthetic vector by Agrobacterium-mediated transformation. InDels formed by CRISPR/Cas9 were detected by sequence analysis. Our results showed that the biosynthesis of BIAs (e.g. morphine, thebaine) was significantly reduced in the transgenic plants suggesting that 4′OMT2 was efficiently knocked-out by our CRISPR-Cas9 genome editing approach. In addition, a novel uncharacterized alkaloid was observed only in CRISPR/Cas9 edited plants. Thus, the applicabilitiy of the CRISPR/Cas9 system was demonstrated for the first time for medicinal aromatic plants by sgRNAs transcribed from both synthetic and viral vectors to regulate BIA metabolism and biosynthesis

Procedure di valutazione dei processi diacronici di modificazione ambientale nelle aree di interesse archeologico da sottoporre a dispositivi di copertura sull'Isola di Vivara

In: La ricerca archeologica a Vivara e le attività dei laboratori dell’Istituto Universitario Suor Orsola Benincasa

Procedure di valutazione dei processi diacronici di modificazione ambientale nelle aree di interesse archeologico da sottoporre a dispositivi di copertura sull’isola di Vivara.

In: La ricerca archeologica a Vivara e le attività dei laboratori dell’Istituto Universitario Suor Orsola Benincasa

Evidence of dramatic biodiversity loss in a wet biotope calls for urgent conservation strategies

The small lake ‘‘Lago di Corree’’, located in the middle Volturno plain (Campania Region, Italy), is a biotope belonging to the Roccamonfina–foce Garigliano Regional Park. It is a ‘‘habitat island’’ in a landscape matrix characterised by intensive agriculture. In the present investigation, plant biodiversity loss, assessed by means of historical and recent field floristic surveys, is related to the lake dynamics and landscape transformation inferred using remote sensing (photointerpretation). A dramatic reduction of plant diversity, and the regional-scale local extinction of several aquatic species are reported

The non-native vascular flora of Campania (southern Italy)

<p>We discuss several aspects of the alien flora of Campania (southern Italy), on the basis of an updated and annotated checklist. The non-native flora includes at present 346 taxa in 231 genera, representing 82 families. However, other 55 taxa were not found later than 1950. Trends in alien plants invasion throughout the investigated area are discussed. Results are therefore compared with similar data from other Italian regions and other European countries. In the Appendix, taxa are listed with essential synonymy, local and general distribution, status, biological form, habitat, diffusion mechanisms, potential threats, literature data, voucher specimens, and miscellaneous ethnobotanical, chronological and taxonomical notes. In addition, 49 escaped taxa are known only for Botanical and private gardens, while 66 taxa are regarded as doubtful aliens. By including also these two categories, the non-native flora of Campania reaches 516 taxa. In addition, numerous doubtful and erroneous indications were discussed. Finally, a reference list for the exotic regional flora from 1500 C.E. to present is proposed.</p