KARAKTERISASI GENETIK ISOLAT Bacillus thuringiensis BERDASARKAN GEN PENANDA 16S rRNA

mega tyas prihatin

Abstract


Bacillus thuringiensis merupakan bakteri berbentuk batang, tidak berkapsul, bersifat gram positif, bersifat motil dengan flagel peritrik, serta membentuk spora. Bakteri ini merupakan kelompok bakteri dalam grup Cereus. Lebih dari 82 serovar Bt diidentitifkasi dengan analisis antigen flagellar (serotyping). Namun, klasifikasi ini memiliki keterbatasan dalam mendeferensiasi kekerabatan spesies ini. Setelah dilakukan uji PCR dengan primer general 16s rRNA dan di sequencing menggunakan sanger sequencing. Sequence DNA kemudian di lakukan analisis menggunakan software MEGA X. Aligment dengan reference yang ada di genebank menggunakan statistik clustal W dan kemudian dihitung jarak genetisnya. Dari hasil perhitungan jarak genetis, isolat Salatiga terbagi menjadi 3 kelompok besar dengan jarak genetis antar isolat 0, yaitu kelompok 1  isolat Salatiga 2,8,11,12,13,17 dan 19. Kemudian kelompok isolat Salatiga 6 dan 7, terakhir isolat salatiga 9.  Kelompok 1 dan 2 memiliki jarak genetis sebesar 0,55%, kelompok 1 dan 3 sebesar 0,68 persen, kelompok 2 dan 3 sebesar 0,14%. Dari perhitungan jarak genetis kemudian di konstruksi menjadi pohon filogeni. Dari konstruksi pohon filogeni, isolat salatiga terbagi menjadi dua klaster. Klaster 1 yaitu isolat salatiga 2,8,11,12,13,17 dan 19 dekat dengan B.thuringiensis serovar israelensis. Sedangkan klaster 2 isolat 6,7 dan 9 dekat dengan B.thuringiensis serovar kurstaki. Dari hasil analisis data sekunder bakteri grup Cereus di genebank dan kontruksi pohon filogeninya. gen 16s rRNA saja tidak cukup untuk membedakan dan mengelompokan grup bakteri Cereus.


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DOI: https://doi.org/10.33992/meditory.v11i1.2310

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