Fadjrianto, Nursandy Yoga and Dr. Ir. Joni Kusnadi, M.Si and Prof. Dr. Ir. Estri Laras Aruminingtyas, M.Sc.St. (2024) Efektivitas Skrining Tingkat Kepedasan Cabai Menggunakan Marka SCAR Berbasis Sekuens RAPD dengan Teknik PCR Singlepleks dan Multipleks. Sarjana thesis, Universitas Brawijaya.
Abstract
Kegemaran masyarakat Indonesia terhadap makanan pedas menyebabkan adanya peningkatan konsumsi cabai di Indonesia setiap tahunnya. Rasa pedas pada cabai tersebut berasal dari senyawa capsaicin. Guna memenuhi kebutuhan mayarakat Indonesia pada komoditas cabai, dilakukanlah pemuliaan tanaman agar dapat memperbaiki kualitas serta kuantitas tanaman cabai. Salah satu tahapan dalam pemuliaan tanaman adalah seleksi. Seleksi dengan metode konvensional memerlukan waktu tunggu yang lama dengan hasil yang tidak selalu konsisten. Seiring dengan perkembangan ilmu bioteknologi, tahapan seleksi dalam pemuliaan tanaman mulai dilakukan dengan pendekatan marka molekuler. Teknik marka molekuler dipilih, karena memiliki beberapa keunggulan dalam hal waktu, usaha, dan keakuratan data dibandingkan dengan metode seleksi secara konvensional. Tujuan dari penelitian ini adalah untuk membandingkan efektivitas primer SCAR-RAPD dalam membedakan antara empat cabai pedas dan empat cabai tidak pedas menggunakan teknik PCR singlepleks dan PCR multipleks . Penelitian akan dilakukan secara deskriptif kualitatif dan kuantitatif. Tahap awal penelitian ini adalah isolasi DNA dari daun tanaman cabai muda yang masih memiliki tekstur lunak dan belum menghasilkan banyak metabolit sekunder. Daun diisolasi menggunakan metode CTAB. Konsentrasi dan kemurnian DNA yang telah diperoleh diukur menggunakan spektrofotometer nanodrop dan diencerkan hingga konsentrasi 100 ng/μl. Setiap sampel DNA kemudian diamplifikasi dengan 1 jenis pasangan primer untuk teknik singlepleks dan dengan semua jenis pasangan primer untuk setiap sampel DNA untuk teknik multipleks.
English Abstract
The Indonesian people's penchant for spicy food has led to an increase in chili consumption in Indonesia every year. The spicy taste of chilies comes from the capsaicin compound. To meet Indonesian people's needs for chili commodities, plant breeding is carried out to improve the quality and quantity of chili plants. One of the stages in plant breeding is selection. Selection using conventional methods requires a long waiting time with inconsistent results. Along with the development of biotechnology, the selection stages in plant breeding began to be carried out using a molecular marker approach. The molecular marker technique was chosen because it has several advantages in terms of time, effort, and data accuracy compared to conventional selection methods. The purpose of this study was to compare the effectiveness of the SCAR-RAPD primer in distinguishing four spicy and four non-spicy chilies using singleplex PCR and multiplex PCR techniques. This research will be carried out in qualitative and quantitative descriptive. The early stage of this research was DNA isolation of young chili leaves which which still have a soft texture and have not produced many secondary metabolites. The young chili leaf was isolated using the CTAB method. The concentration and purity of the DNA that had been obtained were measured using a nanodrop spectrophotometer and diluted to a concentration of 100 ng/μl. Each DNA sample is then amplified with 1 type of primer pair for the singleplex technique and with all types of primer pairs for each DNA sample for the multiplex technique. The amplification results will be visualized using a gel documentary to look at the polymorphic bands. From the research results, it was found that two primer pairs were able to form polymorphic bands in pungent chilies only, namely 24_CR_NK-FR (300,700, dan >1500 bp) and SCAR UBC 145-FR (100–200 bp), a pair of primers that was able to form polymorphic bands in non-pungent chilies only, namely SCAR UBC 145-F with SCAR UBC 20-R (± 350 bp), and a pair of primers that was able to form polymorphic bands both in pungent (± 1000 bp dan >1500bp) or non-pungent (>1500bp) chilies, namely SCAR UBC 145-F with SCAR UBC 20-R. The polymorphic bands that showed indicated the existence of genetic diversity due to the large differences in SHU values between the pungent and non-pungent chili samples used. However, the four pairs of primers were not able to show consistent polymorphic bands when amplified by the multiplex PCR technique, so the singleplex PCR technique was more effective in screening the level of spiciness of chilies based on the polymorphic bands formed
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | 0524100032 |
| Uncontrolled Keywords: | Cabai (Capsicum sp.), PCR Multipleks, PCR Singlepleks, RAPD, SCAR-v |
| Divisions: | Fakultas Teknologi Pertanian > Keteknikan Pertanian |
| Depositing User: | soegeng Moelyono |
| Date Deposited: | 30 Apr 2024 01:36 |
| Last Modified: | 30 Apr 2024 01:36 |
| URI: | http://repository.ub.ac.id/id/eprint/218142 |
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