Penambahan Massa Jamur Semi-kultur untuk Meningkatkan Sensitivitas Deteksi Entomopatogen dengan Metode Quantitative real time Polymerase Chain Reaction

Syaiful Amri Saragih, Wan Arfiani Barus, Widihastuty Widihastuty, Dafni Mawar Tarigan

Abstract


Banyak penelitian telah dilakukan untuk mendeteksi jamur entomopatogen di dalam tanah menggunakan metode qPCR. Namun, beberapa penelitian menunjukkan bahwa metode qPCR belum mampu mendeteksi jamur di sampel tanah yang disebabkan oleh sedikitnya kemelimpahan jamur pada sampel tanah yang diuji. Untuk mengatasi masalah tersebut, pada penelitian ini dilakukan penambahan massa jamur target di dalam tanah pada tingkat tertentu melalui metode semi-kultur dengan tujuan untuk meningkatkan sensitivitas qPCR. Pertama, dipilih media selektif yang cocok untuk kultur keempat target jamur entomopatogen (Cordyceps militaris, Beauveria bassiana, Isaria fumosorosea, dan Metarhizium anisopliae). Kedua, dengan menggunakan media yang dikembangkan, hubungan antara unit pembentuk koloni (CFU) dan DNA tanah standar ditentukan. Ketiga, qPCR dilakukan terhadap ekstrak DNA yang diperoleh dari koloni jamur tiap level. Hasil menunjukkan bahwa media kultur tipe 3b menjadi media selektif yang terbaik untuk kultur jamur. Namun, hubungan linier dengan korelasi kuat antara suspensi jamur dengan nilai Ct belum dapat diperoleh sehingga metodologi semi-kultur tidak berhasil meningkatkan sensitivitas deteksi jamur entomopatogen dari sampel tanah menggunakan metode qPCR. Modifikasi ekstraksi DNA untuk meningkatkan kepadatan DNA dalam ekstrak perlu dilakukan untuk keberhasilan deteksi jamur entomopatogen.


Keywords


Cordyceps militaris, Beauveria bassiana, Isaria fumosorosea, Metarhizium anisopliae.

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References


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DOI: https://doi.org/10.30596/agrium.v27i1.17633

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