NITA ETIKAWATI
Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sebelas Maret. Jl. Ir. Sutami 36A, Surakarta 57126, Central Java, Indonesia

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Primer design of the CO1 gene (Cytochrome Oxidase-1) for Sumatran elephant (Elephas maximus sumatranus) for rapid detection using real-time PCR method MAQQITA TUNJUNG SARI; ARI SUSILOWATI; SETIA BETARIA ARITONANG; OKID PARAMA ASTIRIN; NITA ETIKAWATI; VIRA SAAMIA
Biodiversitas Journal of Biological Diversity Vol. 25 No. 10 (2024)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d251044

Abstract

Abstract. Sari MT, Susilowati A, Aritonang SB, Astirin OP, Etikawati N, Saamia V. 2024. Primer design of the CO1 gene (Cytochrome Oxidase-1) for Sumatran elephant (Elephas maximus sumatranus) for rapid detection using real-time PCR method. Biodiversitas 25: 3840-3849. The population of Sumatran elephants is classified as endangered according to the IUCN due to conflicts between humans and elephants, resulting in many elephants being killed by humans to obtain ivory to be sold. The traded ivory is often processed into other ornaments. A rapid forensic examination with real-time PCR using the molecular approach of the CO1 gene (Cytochrome Oxidase-1) is necessary to determine the original species of the processed elephant ivory. This research aims to design and optimize a primer for identifying Sumatran elephants (Elephas maximus sumatranus). The CO1 gene primer for Sumatran elephants was designed using the Primer3Plus and IDT websites. The primers were used to amplify and identify the species of Sumatran elephants from blood, feces, and urine samples. Blood and feces samples were collected from Sumatran elephants at Taru Jurug Zoo Surakarta, while feces and urine samples were taken from Sumatran elephants at Ragunan Zoo Jakarta. DNA from blood and urine samples was extracted using the TIANamp Genomic DNA Kit, and DNA from feces samples was extracted using the TIANamp Stool DNA Kit. The purity and concentration of the obtained DNA extracts were measured. The designed primers were synthesized by Macrogen (Korea), and primer optimization was performed with a PCR gradient. The identification process of Sumatran elephant DNA extract samples was conducted by amplification using real-time PCR quantification with the standard curve method. The results of this research include the COI-270 primer with the sequences 3'-TTAGGTCAACCAGGCTCTCTTC-5' and 5'-AGGATATACGGTCCAACCAGTG-3', capable of amplifying the target 270 bp and having a single peak at the melt curve temperature of 79°C. The identification protocol with specific COI-270 primer can be used in forensic examinations to identify samples originating from Sumatran elephants.
Gamma-ray irradiation induced polymorphism in Echinacea purpurea revealed by RAPD markers AGUSTINA PUTRI CAHYANINGSIH; NITA ETIKAWATI; AHMAD YUNUS
Nusantara Bioscience Vol. 17 No. 2 (2025)
Publisher : Smujo International

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/nusbiosci/n170211

Abstract

Abstract. Cahyaningsih AP, Etikawati N, Yunus A. 2025. Gamma-ray irradiation induced polymorphism in Echinacea purpurea revealed by RAPD markers. Nusantara Bioscience 17: 289-297. One of the major challenges in the development and cultivation of Echinacea purpurea in Indonesia is the narrow genetic diversity and the lack variation of local accessions, which restricts breeding potential to produce superior varieties with improved traits in terms of morphological characteristics and phytochemical content as medicinal ingredients. To overcome this challenge, gamma-ray irradiation methods can be used as an effective tool to induce mutations and increase genetic variability. This study investigated the impact of gamma irradiation on the genetic diversity of E. purpurea using Random Amplified Polymorphic DNA (RAPD) markers. The seeds of E. purpurea were irradiated using gamma-ray with irradiation doses of 0 Gy (control), 20 Gy, 40 Gy, and 60 Gy. Nine RAPD primers, including OPA-10, OPA-16, OPA-18, and OPA-19, were used to amplify DNA segments. The binary data obtained from the electrophoresis visualization for each treatment were analyzed using the Dice similarity index to calculate the similarity index. Dendrogram construction were performed using the NTSYS. The analysis revealed a significant increase in genetic diversity at doses of 40 Gy and 60 Gy, with 58.04% of the total bands showing polymorphism across all treatments. The 60 Gy treatment, in particular, resulted in the highest genetic dissimilarity compared to the control, indicating a dose-dependent response. The similarity coefficients between control and irradiated plants ranged from 63.7% (20 Gy) to 84.4% (0 Gy), with a noticeable trend toward greater genetic differentiation as the irradiation dose increased. These findings suggest that gamma irradiation effectively induces genetic variation in E. purpurea, which could be harnessed for mutation breeding programs aimed at improving desirable traits such as phytochemical production. However, further studies with larger sample sizes and long-term evaluation are needed to assess the stability of these mutations and their potential for incorporation into breeding programs. This study provides preliminary evidence supporting the use of gamma irradiation in enhancing genetic diversity and its potential application in breeding superior E. purpurea varieties with improved agronomic or medicinal traits.