**3. Results**

### *3.1. Barcode Amplification and Sequencing Success*

The core plant barcode markers rbcL, matK, and ITS2 were amplified at various temperature gradients and sequenced successfully (Figure 2).

**Figure 2.** PCR amplification and sequencing success rate for the DNA barcode markers. (**a**) Annealing temperature gradients used for the barcode amplification, (\* represents the dual annealing temperature 50|55 ◦C), (**b**) PCR amplification and sequencing success of the attempted samples.

The rbcL and ITS2 markers showed significant success rates in PCR amplification ranging from 70% to 95%, while matK exhibited a lower success rate of about 60% (Figure 2). In sequencing, the rbcL marker showed the highest (80%) success rate, followed by the matK (76%) and ITS2 (75%) (Figure 2). In addition, another pair of matK markers was used for the amplicon recovery, where only one sample was successfully amplified. Overall, 99 sequences were obtained belonging to the rbcL (35), matK (34), and ITS2 (30) markers, representing about 13 species.
