CONFIRMATION OF ENDOPHYTIC MICROBES CAUSING CONTAMINATION IN WATER SPINACH (Ipomoea aquatica Forssk.) TISSUE CULTURE
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Abstract
Konfirmasi Mikroba Endofit Penyebab Kontaminasi pada Kultur Jaringan Kangkung (Ipomoea aquatica Forssk.)
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’.
Keywords: Ipomoea aquatica, kultur jaringan, mikroba endofit, morfologi, pohon filogeni
Water spinach in nature lives in symbiosis with endophytic microbes, which have the potential to become contaminants in water spinach tissue culture because they are difficult to eliminate during the explant sterilization process. This study aimed to confirm endophytic microbes that cause contamination in the tissue culture of 'Tetraploid' water spinach so that it can provide initial information for an effective sterilization method. Fourteen contaminant samples in water spinach tissue culture media were isolated and identified molecularly based on the 16S rDNA gene for bacteria, the D1/D2 region of the LSU rRNA gene sequences for yeast, and ITS region of the rDNA gene for mold. The diversity of microbial species identified was compared with the diversity of endophytic microbial types from water spinach plant tissue grown on sterile tissue culture media, microbially contaminated tissue culture media, sterile and non-sterile soil mixed planting media, and water spinach obtained from the market. The results confirmed that endophytic yeast from Ustilaginaceae group (basidiomycetous yeast) derived from water spinach plant tissue was the same type of microbe that contaminated the 'Tetraploid' water spinach tissue culture media. The results gave new information that yeast from the Ustilaginaceae (basidiomycetous yeast) group was the most dominant microbe contaminating water spinach ‘Tetraploid’ tissue culture media. This group is endophytic yeast that lives within Ipomoea aquatica tissues.
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