Source: PlantaPhile^[1]

Source: PlantaPhile^[2]

Saw Palmetto (Serenoa repens)

Amplification primers:

matK mini–barcode forward (5′-TCT AAA GCT AAA TTT TGT ACC GTA TCG-3′) and reverse (5′-GCC AAA GTT CTA GCA CAC GAA-3′)

rbcL mini–barcode forward (5′-GCT ATC ACA TCG AAA CCG TTG-3′) and reverse (5′-GCC TTG GAA AGT TTT GGA ATA AG-3′)

PCR reaction mixture:

matK mini–barcode: 15 µL final volume containing 1.5 µL PCR buffer (200 mM tris [pH 8.8], 100 mM KCl, 100 mM (NH₄)₂SO₄, 20 mM MgSO₄, 1% [v/v] Triton X-100, 50% [w/v] sucrose, 0.25% [w/v] cresol red), 0.2 µM dNTPs, 0.025 μg/μL bovine serum albumin (BSA), 1 μM of each primer, 0.25 units of Taq polymerase, and 0.5 µL template DNA.

rbcL mini–barcode: 15 µL final volume containing 1.5 µL PCR buffer (200 mM tris [pH 8.8], 100 mM KCl, 100 mM (NH₄)₂SO₄, 20 mM MgSO₄, 1% [v/v] Triton X-100, 50% [w/v] sucrose, 0.25% [w/v] cresol red), 0.2 µM dNTPs, 0.025 μg/μL bovine serum albumin (BSA), 48 mM betaine, 1 μM of each primer, 0.25 units of Taq polymerase, and 0.5 µL template DNA.

PCR program: 95°C for 2.5 min; 35 cycles: 95°C for 30 s, 60°C for 30 s; 60°C for 10 min.

Sequencing primers: Same as amplification primers.

Sequence:

matK mini–barcode: 5′-GGG CAT CCT ATT AGT AAG CCG ATC TGG ACC GAT TTA TCA GAT TGC GAT ATT ATT GAT CGA TTT GGT CGG ATA TGT AGA AAT CTT TCT CAT TAT CAC AGT GGA TCC TCA AAA AAA CAG AGT TTG TAT CGA ATA AAG TAT ATA CTT CGG TT-3′

rbcL mini–barcode: 5′-TCG GGG AGG AAA ATC AAT ATA TTG CTT ATG TAG CTT ATC CTT TAG ACC TTT TTG AAG AAG GTT CTG TTA CTA ACA TGT TTA CTT CCA TTG TGG GTA ATG TAT TTG GTT TCA AAG CCC TAC GAG CTC TAC GTC TGG AGG ATC TGC GAA TTC CCA CTT-3′

Diagnostic positions:

matK mini–barcode

                                  66667777777778888
                                  78893455668890111
                                  85958437565682678
Brahea                            YGGCCTCCCAACCTCGA
Rhapidinae Chamaerops             C..Y..YY....Y....
Rhapidinae Guihaia                C................
Rhapidinae Maxburretia            C................
Rhapidinae Rhapidophyllum         C................
Rhapidinae Rhapis                 C................
Rhapidinae Trachycarpus           C................
Livistoninae Johannesteijsmannia  C....W...RR......
Livistoninae Lanonia              CR...............
Livistoninae Licuala              CR..Y...Y.....M..
Livistoninae Livistona            C................
Livistoninae Pholidocarpus        C..........Y.A..R
Livistoninae Pritchardiopsis      C..............T.
Livistoninae Saribus              CRK............K.
Acoelorrhaphe wrightii            C...............G
Serenoa repens                    C...............G

rbcL mini–barcode

                                  222233
                                  558909
                                  130218
Brahea                            GGTCAA
Rhapidinae Chamaerops             ......
Rhapidinae Guihaia                ......
Rhapidinae Maxburretia            ......
Rhapidinae Rhapidophyllum         ......
Rhapidinae Rhapis                 ..C...
Rhapidinae Trachycarpus           ......
Livistoninae Johannesteijsmannia  .....C
Livistoninae Lanonia              ??????
Livistoninae Licuala              RR...M
Livistoninae Livistona            .....M
Livistoninae Pholidocarpus        ....GC
Livistoninae Pritchardiopsis      .....C
Livistoninae Saribus              .....C
Acoelorrhaphe wrightii            ...T.C
Serenoa repens                    ......