CONFIRMATION OF ENDOPHYTIC MICROBES CAUSING CONTAMINATION IN WATER SPINACH (Ipomoea aquatica Forssk.) TISSUE CULTURE
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Abstract
Tanaman kangkung secara alami bersimbiosis dengan mikroba endofit, yang berpotensi menjadi kontaminan pada kultur jaringan kangkung karena berada di dalam jaringan dan sulit dijangkau saat proses sterilisasi eksplan. Tujuan penelitian ini adalah mengonfirmasi mikroba endofit penyebab kontaminasi pada kultur jaringan kangkung ‘Tetraploid’ sehingga dapat menjadi informasi awal untuk metode sterilisasi yang efektif. Sebanyak 14 sampel kontaminan pada media tanam kultur jaringan kangkung diisolasi dan diidentifikasi secara molekuler berdasarkan gen 16S rDNA untuk bakteri, daerah D1/D2 dari gen LSU rRNA untuk khamir, dan daerah ITS dari gen rDNA untuk jamur. Keragaman jenis mikroba yang teridentifikasi dibandingkan dengan keragaman jenis mikroba endofit dari jaringan tanaman kangkung yang ditanam pada media kultur jaringan tidak terkontaminasi mikroba, media kultur jaringan terkontaminasi mikroba, media tanam campuran tanah steril dan tidak steril, serta kangkung yang didapatkan dari pasar. Hasil penelitian ini mengonfirmasi bahwa khamir endofit dari kelompok Ustilaginaceae (basidiomycetous yeast) yang berasal dari jaringan tanaman kangkung sama dengan jenis kontaminan yang mengontaminasi media kultur jaringan kangkung ‘Tetraploid’. Khamir dari kelompok Ustilaginaceae (basidiomycetous yeast) tersebut juga merupakan mikroba paling dominan yang mengontaminasi media tanam kultur jaringan kangkung ‘Tetraploid’.
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