Indaryani, Patricia Martina Kusuma (2019) Analisis Keragaman Genetik Tanaman Anggur (Vitis vinivera L.) Varietas BS 60 Hasil Perbanyakan Secara Kultur Jaringan Dengan Marka ISSR (Inter Simple Sequence Repeats). Sarjana thesis, Universitas Brawijaya.
Abstract
Tujuan konservasi adalah melestarikan plasma nutfah agar dapat dimanfaatkan untuk masa kini dan mendatang. Dalam kegiatan tersebut, pemeliharaan stabilitas genetik merupakan hal yang sangat penting untuk menjamin plasma nutfah yang disimpan tetap sesuai dengan identitas asalnya. Keragaman genetik dapat diketahui lebih awal melalui karakterisasi molekuler. Karakterisasi molekuler memiliki keunggulan, yaitu waktu yang dibutuhkan lebih cepat karena tidak perlu menunggu tanaman hingga fase dewasa, mampu membedakan individu berkerabat dekat, dan tidak dipengaruhi oleh faktor lingkungan. Inter Simple Sequence Repeat (ISSR) merupakan penanda molekuler yang umumunya digunakan untuk studi keanekaragaman genetika, filogeni, penandaan gen, pemetaan genom dan biologi evolusi pada berbagai tanaman. Seperti tanaman yang lain, tanaman anggur dapat diperbanyak secara kultur jaringan. Perbanyakan secara kultur jaringan tidak menutup kemungkinan terjadinya variasi somaklonal yang menyebabkan variasi atau keanekaragaman pada tanaman hasil perbanyakan kultur jaringan. Maka dari itu diperlukan analisis keragaman genetik tanaman anggur varietas BS 60 hasil perbanyakan secara kultur jaringan agar mengetahui tingkat keanekaragaman yang terjadi. Penelitian dilaksanakan pada bulan Januari hingga April 2019 di Balai Penelitian Tanaman Jeruk dan Buah Subtropika yang beralamat di Jalan Raya Tlekung No. 1, Beji, Kecamatan Junrejo, Kota Batu, Provinsi Jawa Timur 65327. Penelitian akan dilakukan dengan menggunakan metode deskriptif dengan penggunaan sampel berdasarkan 3 jenis kategori antara lain, sampel yang disubkultur 14 kali, sampel disubkultur 15 kali, sampel tanpa masa kultur dengan perbanyakan secara vegetatif. Masing – masing kategori menggunakan 10 sampel tanaman untuk di ekstraksi, uji kualitas DNA, analisis Polymerase Chain Reaction (PCR) hingga elektroforesis DNA. Tahap analisis Polymerase Chain Reaction (PCR) menggunakan 5 jenis primer ISSR. Pengamatan dilakukan terhadap profil pita DNA yang dihasilkan melalui proses elektroforesis. Penambahan atau kehilangan pita DNA dibandingkan dengan kontrol menunjukkan adanya off-type. Masing – masing pita DNA diperlakukan sebagai sebuah unit karakter. Pita DNA yang terbentuk diberi skor untuk pita terbentuk (1) atau pita tidak terbentuk (0) yang selanjutnya menjadi sebuah matriks biner. Data kemudian dianalis dengan analisis kluster unweighted pair group method arithmetic averages (UPGMA) dan divisualisasikan dengan menggunakan program NTSYS PC versi 2.20. Berdasarkan hasil penelitian dapat diketahui bahwa penanda molekuler ISSR dapat mendeteksi keragaman anggur yang dikonservasi secara kultur jaringan. Penanda molekuler ISSR mampu menghasilkan jumlah total pita polimorfisme yang dihasilkan pada kelima primer sebanyak 34 buah, sedangkan jumlah total pita monomorfisme yang dihasilkan pada kelima primer sebanyak 30 buah. Persentase polimorfisme tertinggi terdapat pada primer L8 sebesar 66,6%. Persentase polimorfisme terendah terdapat pada primer L9 sebesar 33,3 %. Berdasarkan hasil analisis kluster unweighted pair group method arithmetic averages (UPGMA), diperoleh dendogram keragaman genetik tanaman anggur dengan tingkat keragaman genetik tanaman anggur berkisar antara 66 – 100% atau jarak genetik berkisar antara 0 – 34%.
English Abstract
The purpose of conservation is to preserve germplasm so that it can be used for the present and future. In these activities, maintaining genetic stability is very important to ensure the stored germplasm remains in accordance with its true identity. Genetic diversity can be known earlier through molecular characterization. Molecular characterization has the advantage, that it takes a shorter time due to unnecessary waiting for crops to mature phase, is able to diferentiate between closely related individuals, and not influenced by environmental factors. Inter Simple Sequence Repeat (ISSR) is a molecular marker commonly used for studies of genetic diversity, phylogeny, gene marking, genome mapping and evolutionary biology in various plants. Like other plants, grape can be propagated by tissue culture. Propagation by tissue culture that has the possibility of somaclonal variation that causes variation or diversity in plants produced by multiplication of tissue culture. Therefore it is necessary to analyze the genetic diversity of BS 60 grape vines as a result of tissue culture propagation in order to determine the level of diversity. The research was conducted in January 2019 until April 2019 at the Research Institute for Citrus and Subtropical Fruits located at Tlekung Street No 1, Beji, District of Junrejo, Kota Batu, East Java Province 65327. The study was conducted by using descriptive method with the use of samples based on three types of categories, the samples were subcultured 14 times, samples were subcultured 15 times, samples without the culture period with vegetative propagation. Each category uses 10 plant samples for extraction, DNA quality testing, polymerase chain reaction (PCR) and DNA electrophoresis. The phase of polymerase chain reaction (PCR) uses 5 types of primary ISSR. Observations based on the DNA band profile based on electrophoresis results. Addition or loss of DNA bands compared to controls showed an off-type. Each DNA band is treated as a character unit. The formed DNA tape is given a score for the formed band (1) or the band is not formed (0) which then becomes a binary matrix. Data was then analyzed by analyzing cluster unweighted pair group method arithmetic averages (UPGMA) and visualized using program NTSYS PC version 2.20. Based on the results of the study, molecular markers of ISSR can detect the genetic diversity of grapes conserved by tissue culture. Molecular markers of ISSR were able to produce the total number of polymorphism bands produced in all five primers as many as 34 bands, while the total number of monomorphism bands produced in the five primers was 30 bands. The highest percentage of polymorphism is found in L8 primers of 66.6%. The lowest percentage of polymorphism is found in L9 primers of 33.3%. Based on the results of cluster analysis of unweighted pair group method arithmetic averages (UPGMA), obtained dendogram of genetic diversity of grape with a degree of genetic diversity of grape ranging from 66 - 100% or genetic distance ranging from 0 - 34%.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | SKR/FP/2019/253/051906996 |
| Uncontrolled Keywords: | - |
| Subjects: | 600 Technology (Applied sciences) > 634 Orchards, fruits, forestry > 634.8 Grapes / Viticulture |
| Divisions: | Fakultas Pertanian > Agroekoteknologi |
| Depositing User: | soegeng sugeng |
| Date Deposited: | 24 Aug 2020 07:05 |
| Last Modified: | 24 Aug 2020 07:05 |
| URI: | http://repository.ub.ac.id/id/eprint/173253 |
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