TRANSFORMASI GEN PADA NILAM UNTUK KETAHANAN TERHADAP PENYAKIT UTAMA MENGGUNAKAN Agrobacterium tumefaciens

NFN Sukamto, Tri Joko Santoso, Atmitri Siharmini, Aniversari Apriana, NFN Amalia, Nursalam Sirait

Abstract


Nilam banyak dibudidayakan di Indonesia, dan lebih dari 80% produksi minyak nilam dunia dipasok dari Indonesia. Masalah utama dalam budidaya nilam di Indonesia adalah penyakit, seperti penyakit layu bakteri, budok dan nematoda. Sampai saat ini varietas tahan terhadap penyakit, khususnya budok, belum diperoleh. Tanaman tahan dapat diperoleh dengan teknik transformasi gen. Transkripsi faktor WRKY telah diketahui dapat meregulasi serangan beberapa patogen penyebab penyakit tanaman. Gen OsWRKY76 terletak pada segmen kromosom 9 tanaman padi yang telah diidentifikasi terkait dengan ketahanan berspektrum luas. Penelitian bertujuan untuk mengintroduksikan konstruksi gen OsWRKY76 yang berasal dari padi ke dalam tanaman nilam melalui bantuan Agrobacterium tumefaciens. Pada percobaan pertama, tanaman nilam ditransformasi dengan A. tumefaciens strain EHA 105 yang mengandung gen OsWRKY76. Perlakuan terdiri atas waktu induksi eksplan yang akan ditransformasi (pre-kultur) di dalam medium MS, yaitu 5 dan 7 hari, dan waktu inokulasi A. tumefaciens yaitu 10 dan 20 menit. Pada percobaan kedua, analisis molekuler untuk mengkonfirmasi keberadaan gen OsWRKY76 dalam tanaman nilam menggunakan teknik PCR dengan primer hptII. Hasil penelitian menunjukkan waktu induksi eksplan terbaik sebelum transformasi adalah 5 hari, dengan perendaman di dalam suspensi A. tumefaciens selama 10 menit. Dari transformasi tersebut telah dihasilkan 187 kalus independen. Hasil analisis PCR terhadap galur-galur putatif transgenik independen, lima galur (T1, T8, T10, T11, T13) positif mengandung gen hptII, yaitu gen penanda ketahanan terhadap antibiotik higromisin yang berada satu konstruk dengan gen OsWRKY76. Hasil tersebut menunjukkan bahwa gen OsWRKY76 yang berasal dari padi dapat diintroduksikan pada tanaman nilam dan berpeluang sebagai kandidat tahan terhadap penyakit utama.


Keywords


Pogostemon cablin Benth; gen OsWRKY76; ketahanan terhadap penyakit.

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DOI: http://dx.doi.org/10.21082/bullittro.v28n1.2017.37-46

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