Keragaman Jumlah Salinan Transgen Galur T0 Padi Kultivar Nipponbare Berdasarkan Analisis qPCR dengan Penanda Gen hptII

Aqwin Polosoro, Wening Enggarini

Abstract


Keragaman Jumlah Salinan Transgen CsNitr1-L pada Galur Transforman T0 Padi Kultivar Nipponbare. Perakitan tanaman transgenik dengan menggunakan bantuan Agrobacterium tumefaciens menghasilkan penyisipan transgen yang berbeda, baik dalam jumlah salinan maupun letak transgen dalam genom tanaman. Penelitian ini bertujuan untuk menganalisis keberadaan kimera dan jumlah salinan pada tiap anakan dalam satu rumpun dan beberapa rumpun padi Nipponbare transforman T0 yang berasal dari kalus yang sama. Gen CsNitr1-L pada plasmid biner pCAMBIA1300 ditranformasikan ke dalam genom tanaman padi kultivar Nipponbare dengan menggunakan A. tumefaciens strain LBA 4404. Analisis molekuler dilakukan terhadap tiga anakan dari masing-masing empat rumpun tanaman Nipponbare transgenik generasi T0 (event 1, 2, 3, dan 4) dan empat kelompok rumpun tanaman T0 yang berasal dari kalus yang sama. Dari masing-masing kelompok tersebut diambil 3 rumpun T0. Hasil analisis qPCR menunjukkan bahwa anakan-anakan yang berasal dari satu rumpun tanaman T0 memiliki jumlah salinan transgen yang seragam. Selain itu, hasil analisis qPCR juga mengungkap bahwa tidak semua tanaman yang berasal dari kalus yang sama memiliki jumlah salinan transgen yang seragam. Implikasi dari hasil ini bahwa setiap rumpun padi T0 yang tumbuh dari kalus hasil transformasi perlu dipisah saat aklimatisasi supaya benih T1 yang dihasilkan seragam.  


Keywords


jumlah salinan; qPCR; Agrobacterium tumefaciens; Nipponbare

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DOI: http://dx.doi.org/10.21082/jbio.v12n2.2016.p73-80

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