Fig. 1: Constitutive AMP and fAMP expression from the tobacco plastid genome.
From: Expression strategies for the efficient synthesis of antimicrobial peptides in plastids
a Sequences encoding AMPs or fAMPs were placed under the control of the Nicotiana tabacum plastid ribosomal RNA operon promoter (NtPrrn), the T7 phage gene10 leader (T7g10L) as 5′ UTR and the 3′ UTR of the atpA gene from Chlamydomonas reinhardtii (CrTatpA). The spectinomycin resistance gene aadA for selection of transplastomic lines was driven by the Chlamydomonas psaA promoter and 5′ UTR (CrPpsaA), and the 3′ UTR of the Chlamydomonas atpB gene (CrTatpB). Different combinations of AMPs were linked with linkers of 5 or 15 amino acids (pink). The constructs were flanked by homology regions for integration into the spacer between the trnfM and trnG genes in the tobacco plastid genome. Restriction sites (BglII) and the binding site of the probe for RFLP analysis to verify the transplastomic state are indicated. For information on AMPs and fAMPs, see Supplementary Tables 1, 2 and Supplementary Data 1 and 2. b Seed assays to confirm homoplasmy of transplastomic AMP-expressing lines. Seeds were sown on medium with (Spec+) or without (Spec−) spectinomycin, and photos were taken after two weeks. In the case of 3‐Molluxubin‐5 (3‐M‐5), 3‐Molluxubin‐15 (3‐M‐15) and 3‐Novescandin‐5 (3‐N‐5), seeds were obtained from grafted plants. 3‐N‐15: 3‐Novescandin‐15.
