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USAHA PENURUNAN SERAPAN KADMIUM PADA PADI ‘CIHERANG’ MELALUI PENGEDITAN GEN OsNRAMP5 MENGGUNAKAN TRANSFORMASI METODE BULUH SERBUK SARI

Wulandari, Rani Agustina ; Wulandari, Rani Agustina ; Wulandari, Rani Agustina ; Basunanda, Panjisakti ; Basunanda, Panjisakti ; Basunanda, Panjisakti ; Trijatmiko, Kurniawan Rudi ; Trijatmiko, Kurniawan Rudi ; Trijatmiko, Kurniawan Rudi ; Susanti, Pridana Intan ; Susanti, Pridana Intan ; Susanti, Pridana Intan

Abstract:ABSTRACT Contamination of heavy metal cadmium in rice fields is one of the problems in rice production. OsNRAMP5 is the gene responsible for reducing the absorption of metal Cd. The purpose of this research was to obtain a CRISPR/Cas-9 construct for decrease uptake heavy metal of cadmium in rice and to obtain mutants of T0 generation that mutated in the target gene for decrease in heavy metal uptake of cadmium. The research was carried out at the Laboratory of Molecular Biology and Greenhouse of Research and Development of Biotechnology and Agricultural Genetic Resources (BB BIOGEN) Bogor, West Java from March to August 2017. This research was conducted through two stages: a) construction vector cassette CRISPR/Cas-9 for editing of OsNRAMP5 gene and b) introduction of CRISPR-Cas construct OsNRAMP5 gene to ‘Ciherang’ rice for function validation through pollen tube transformation method. In this research, the gRNA constructs for OsNRAMP5 to decrease in heavy metal cadmium uptake in rice were successfully inserted into the pENTR4:gRNA4 plasmid. However, mutant strains of mutated T0 rice in the target gene for the decrease in heavy metal uptake of cadmium have not been obtained. Keyword: OsNRAMP5, CRISPR/Cas-9, introduction

Intisari:INTISARI Kontaminasi logam berat kadmium di sawah merupakan salah satu masalah yang dihadapi dalam produksi tanaman padi. Gen yang bertanggung jawab untuk mengurangi penyerapan logam Cd, yaitu OsNRAMP5. Tujuan dari penelitian ini adalah mendapatkan konstruk CRISPR/Cas-9 untuk penurunan serapan logam berat kadmium pada padi dan mendapatkan galur-galur mutan padi generasi T0 yang termutasi pada gen target untuk penurunan serapan logam berat kadmium. Penelitian dilaksanakan di Laboratorium Biologi Molekuler dan Rumah Kaca Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetika Pertanian (BB BIOGEN) Bogor, Jawa Barat pada bulan Maret hingga Agustus 2017. Penelitian ini dilakukan melalui dua tahapan, yaitu: a) konstruksi kaset vektor CRISPR/Cas-9 untuk pengeditan gen OsNRAMP5 dan b) introduksi konstruk CRISPR-Cas gen OsNRAMP5 ke tanaman padi kultivar ‘Ciherang’ untuk validasi fungsi melalui transformasi menggunakan metode buluh serbuk sari. Pada penelitian ini, konstruk gRNA bagi OsNRAMP5 untuk penurunan serapan logam berat kadmium pada padi berhasil disisipkan ke dalam plasmid pENTR4:gRNA4. Akan tetapi, galur-galur mutan padi generasi T0 yang termutasi pada gen target untuk penurunan serapan logam berat kadmium belum berhasil diperoleh. Kata kunci: OsNRAMP5, CRISPR/Cas-9, introduksi ===ABSTRACT Contamination of heavy metal cadmium in rice fields is one of the problems in rice production. OsNRAMP5 is the gene responsible for reducing the absorption of metal Cd. The purpose of this research was to obtain a CRISPR/Cas-9 construct for decrease uptake heavy metal of cadmium in rice and to obtain mutants of T0 generation that mutated in the target gene for decrease in heavy metal uptake of cadmium. The research was carried out at the Laboratory of Molecular Biology and Greenhouse of Research and Development of Biotechnology and Agricultural Genetic Resources (BB BIOGEN) Bogor, West Java from March to August 2017. This research was conducted through two stages: a) construction vector cassette CRISPR/Cas-9 for editing of OsNRAMP5 gene and b) introduction of CRISPR-Cas construct OsNRAMP5 gene to ‘Ciherang’ rice for function validation through pollen tube transformation method. In this research, the gRNA constructs for OsNRAMP5 to decrease in heavy metal cadmium uptake in rice were successfully inserted into the pENTR4:gRNA4 plasmid. However, mutant strains of mutated T0 rice in the target gene for the decrease in heavy metal uptake of cadmium have not been obtained. Keyword: OsNRAMP5, CRISPR/Cas-9, introduction

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Penerbit : Departemen Budidaya Pertanian Fakultas Pertanian UGM, 2018
Tahun 2018
Deskripsi viii,56 hlm .; ill .: 12 cm
No Klasifikasi
Bahasa Indonesian
Jenis Penelitian Skripsi
Subyek introduksi
Kata Kunci OsNRAMP5, CRISPR/Cas-9, introduksi
Jumlah Eksemplar 1
Barcode No. Inventaris Lokasi
  fpn   181144FPN   Perpustakaan Fakultas Pertanian