Analisis Sebagian Sekuen Gen Ferritin2 pada Padi (Oryza sativa L.) Indragiri Hilir, Riau

Fadel Nugraha, Dewi Indriyani Roslim, Yolla Putri Ardilla, Herman -

Abstract


Ion Fe bebas sangat beracun bagi tanaman, karena dapat membentuk radikal bebas di dalam sel. Walaupun demikian, tanaman memiliki mekanisme untuk mempertahankan homeostasis Fe di dalam sel yang melibatkan protein ferritin. Penelitian ini bertujuan untuk menganalisis dan membandingkan sekuen nukleotida gen ferritin2 dari dua genotipe padi lokal (Bakung dan Serei) dari Indragiri Hilir, Riau dengan varietas padi rawa unggul tahan cekaman kelebihan Fe (Siam Sintanur), padi unggul tidak tahan cekaman kelebihan Fe (IR64), dan data sekuen nukleotida kultivar Nipponbare. Metode penelitian meliputi isolasi DNA toal dari daun segar tanaman padi menggunakan metode CTAB dan amplifikasi DNA (PCR) menggunakan primer OsFer_F3 (forward) dan Gross_R (reverse). Produk PCR kemudian disekuensing dan disejajarkan. Pada penelitian ini telah diperoleh fragmen dari gen ferritin2 dari kelima genotype atau varietas padi yang diuji, yang berukuran sekitar 1200 pb. Analisis pensejajaran menunjukkan terdapat 56 SNP (single nucleotide polymorphism) pada sekuen nukleotida gen ferritin2 tersebut. Bakung menunjukkan kedekatan yang tinggi dengan Nipponbare, diikuti dengan IR64, Siam Sintanur, dan Serei. Kemungkinan Bakung merupakan genotipe padi lokal dari Indragiri Hilir, Riau yang tahan cekaman kelebihan Fe.

Free Fe ions are highly toxic to plants, because it can form free radicals in the cells. However, plants have mechanisms to maintain Fe homeostasis in the cells involving ferritin proteins. This study was aimed to analyze and to compare the nucleotide sequence of ferritin2 gene in two local rice genotypes (namely Bakung and Serei) from Indragiri Hilir, Riau and in Fe overload-tolerant rice variety (Siam Sintanur), Fe overload-sensitive rice variety (IR64), as well as the nucleotide sequence of Nipponbare rice cultivar. The research methods consisted of DNA isolation from fresh leaves of rice plants using CTAB method and DNA amplification (PCR) using a couple of primers, OsFer_F3 (forward) and Gross_R (reverse). The PCR products were then sequenced and aligned. DNA fragments of ferritin2 gene with length of approx. 1200 bp were obtained from those four rice varieties or genotypes tested. Alignment revealed 56 SNPs (single nucleotide polymorphisms) in the ferritin2 gene sequences. Bakung showed close distance with Nipponbare, followed by IR64, Siam Sintanur, and Serei. It wa suggested that Bakung was Fe overload-tolerant local rice genotype from Indragiri Hilir, Riau.


Keywords


ferritin2 gene; Fe; Oryza sativa; Indragiri Hilir; Riau

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DOI: https://doi.org/10.15294/biosaintifika.v6i2.3102

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