Optimization of primer and polymerase chain reaction conditions to amplify COI locus for identification of Purnajiwa (Euchresta horsfieldii (Lesch.) Benn.) collected from Bedugul, Bali

Abstract

The polymerase chain reaction (PCR) has been used for molecular research to amplify DNA fragments, especially from a small amount of genetic material. PCR has been applied for numerous researches, including for plant molecular identification. A good PCR product is dependent on good amplification of the DNA segment in optimum condition. This research was conducted to optimize the primer combination, and the annealing temperature for DNA barcoding application of Balinese rare medicinal plant (Euchresta horsfieldii (Lesch.) Benn.) collected from Bedugul. COI is a suitable primer to identify unknown species to higher taxa levels and species with high phenotypic plasticity. The range of annealing temperatures was tested to amplify the combination of five COI primers: GWSF and GWSF5 for forwarding primers and GWSR, GWSR3, and GWSR5 for reverse primers. The annealing temperature was 52, 54, 56, 58, 60°C for 30 seconds. The result showed that GWSF-GWSR, GWSF-GWSR5, GWSF5-GWSR, GWSF5-GWSR5, and GWSF5-GWSR3 produced multiple bands, the double band that cannot be cut, multiple bands, faint amplification band for GWSF5-GWSR5 and GWSF5-GWSR3, respectively. Primer combination of GWSF-GWSR3 with an annealing temperature of 52°Cproduced double bands that were available to have proceeded for sequencing. In conclusion, the combination of GWSF-GWSR3 that is annealed at 52°C produced the best amplification band. The primer combinations and conditions can be used for further identification of Purnajiwa collected from Bedugul.

Phytochemical Profiling of Balinese Alkaloid-Source Plant Purnajiwa (Kopsia arborea Blume. and Euchresta horsfieldii (Lesch.) Benn.)

“…Numerous studies have been conducted to determine the molecular identity of this plant and assess its antioxidant activity. [3][4][5][6][7][8] © 2024 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License…”

“…4 Research on K. arborea obtained from Denpasar has been conducted to identify the content of its phytochemical compounds, antioxidants and aphrodisiac activities and ascertain its genomic and taxonomic profile. [4][5][6][7][8] Kopsia arborea Blume is a species in the Kopsia (family Apocynaceae) genus. The genus Kopsia consists of about 30 species distributed in several countries, mainly Asia, China, Australia, and some islands in the Western Pacific.…”

Application of DNA Barcoding for authentication of Balinese traditional medicinal plant Purnajiwa (Kopsia arborea Blume. and Euchresta horsfieldii) (Lesch.) Benn

“…Morphological identification revealed several differences between the purnajiwa. 1 Some studies mention the pranajiwa as Kopsia arborea Blume. or Euchresta horsfieldii (Lesch.)…”

“…[3][4][5] Despite its beneficial phytochemical and pharmacological activities, purnajiwa is considered a rare and endangered plant. 1,2 Genetic studies are an excellent starting point for conservation efforts. Genetic information provides many benefits in aquaculture management and conservation, especially for species with limited numbers and facing extinction.…”