The complete plastome of Nonea vesicaria (L.) Rchb. (Boraginaceae), the first chloroplast genome belonging to the Nonea genus

Abstract The predominantly Western Mediterranean weed Nonea vesicaria (L.) Rchb. can be found in agricultural or other man-made environments. Despite containing some beneficial compounds, extracts from this plant have also been described as detrimental and should be carefully monitored. In this study, the complete chloroplast of N. vesicaria isolate BPTPS250 is described, being the first available plastome from an isolate belonging to the Nonea genus. The chloroplast genome is 151,099 bp in length with a 37.3% GC content. It displays a quadripartite structure that contains a pair of inverted repeat regions (27,012 bp) that separate a large single-copy region (80,041 bp) and a small single-copy region (17,034 bp). A total of 134 genes were predicted, including 89 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The phylogenetic analysis confirmed the placement of N. vesicaria under the Boraginaceae family, belonging to the Boraginales order, with a close relationship with Borago officinalis L. This study will contribute to conservation, phylogenetic, and evolutionary studies, as well as DNA barcoding applications for food and feed safety and quality.

Nonea Medik. (monkswort) is a genus of herbaceous flowering plants (angiosperms) of the Boraginoideae subfamily, part of the Boraginaceae family, that encompasses ca. 42 genera with large flowers, with faucal and basal scales, gynobasic style, and often incurved roughly ovoidal nutlets (Chac on et al. 2016). Nonea includes ca. 45 species distributed in North Africa and from the Temperate Eurasia to the Indian Subcontinent. Although the Nonea centers of diversity are the Pontic-Caucasian and Irano-Turanic mountains, about 35 species occur in the Mediterranean basin, of which nine are found in its European part (WFO 2022).
Nonea vesicaria (L.) Rchb. (Reichenbach 1832) (red monkswort) is an annual or biennial greyish-green hairy weed found in Western Mediterranean's agricultural and other man-made environments. Its natural habitat is the Iberian Peninsula, Balearic Islands, and Sicily, and it is considered a sporadic alien in the U.S.A. and Canada (GBIF Secretariat 2022). Nonea vesicaria has been reported as beneficial in the phytoremediation of oil-polluted soils (Panchenko et al. 2022). Despite harboring stearidonic acid-rich seeds (an omega-3 fatty acid) and being a source of gamma-linolenic acid used for premenstrual syndrome, arthritis, blood pressure, and skin disorders, N. vesicaria extracts have been described as having cytotoxic, hemolytic, and antioxidant activities (Mouffouk et al. 2020), and as such should be carefully monitored.
Total genomic DNA was extracted from young leaves, frozen in liquid nitrogen immediately after collection and kept at À80 C, using an adaptation of the Doyle and Doyle (1987) methodology. The obtained DNA was sheared by sonication using a Bioruptor (Diagenode), libraries were prepared with the NEBNext Ultra II DNA Library Prep Kit (New England Biolabs), and 150 bp paired-end sequencing was performed on an Illumina MiSeq platform using a v2 chemistry kit.
The chloroplast genome of N. vesicaria isolate BPTPS250 (GenBank accession number: OL335187; Supplemental material Figure S3) is 151,099 bp in length with a 37.3% GC content, displaying a quadripartite structure that contains a pair of inverted repeat (IR) regions (27,012 bp, GC content 42.5%), separated by a large single-copy (LSC) region (80,041 bp, GC content 35.2%) and a small single-copy (SSC) region (17,034 bp, GC content 30.9%). A total of 134 genes were predicted, including 37 tRNA genes, 8 rRNA genes, and 89 protein-coding genes.
The phylogenetic analysis (see Supplemental material for additional details) was performed using the concatenated sequences coding for the shared proteome (25 coding sequences) extracted from all 12 verified and complete chloroplast genomes belonging to the Boraginales order available in GenBank (Accession date: 2022-01-10) and from the complete chloroplast genome of N. vesicaria obtained in this study. The sequences were aligned using MAFFT v7.450 (Katoh and Standley 2013) and further analyzed with the IQ-TREE 2 software package (Minh et al. 2020). The best-fit substitution model (TVM þ FþR2 chosen according to the Bayesian Information Criterion) was selected according to ModelFinder (Kalyaanamoorthy et al. 2017), followed by a tree reconstruction (Figure 1) using IQ-TREE (Nguyen et al. 2015) using ultrafast bootstrap with UFBoot (10,000 replicates) (Hoang et al. 2018). The outgroup was Salvia officinalis L. (NC_038165) from the Lamiaceae family belonging to the Lamiales order.
The maximum likelihood tree showed that N. vesicaria is placed under the Boraginaceae family, belonging to the Boraginales order, and has an unsurprising close relationship with Borago officinalis L. (Chac on et al. 2016). Both are likely to belong to the same Boragineae tribe as sister clades, with 100/ 100 per cent support (SH-aLRT/UFBoot2). The phylogenetic analysis performed with the alignments of the Boraginales complete chloroplast genomes also supports the same relationship between N. vesicaria and B. officinalis.
This complete chloroplast genome will contribute to conservation, phylogenetic, and evolutionary studies. It will also support DNA barcoding applications for food and feed safety and quality purposes that target the detection of species with cytotoxic and hemolytic potential.

Author contributions
The authors had the following contribution to the paper: FBGconception and design; JCcollection and taxonomic identification of the studied specimen; ICL and FBGanalysis and interpretation of the data; FBGdrafting of the paper; ICL, JC, MTBC, and FBGcritical revision for intellectual content; FBGfinal approval of the version to be published. All authors agree to be accountable for all aspects of the work herein presented.

Ethical approval statement
The species described and studied in this manuscript is not under legal protection status, either by national or European Union legislation, namely the 92/43/CEE Directive. A careful nondestructive collection protocol for voucher sampling was followed to guarantee the full future reproductive viability of the studied plant population.

Disclosure statement
No potential competing or conflict of interests was reported by the author(s).   Data availability statement