Deteksi Molekuler Burkholderia glumae, Penyebab Penyakit Hawar Malai Padi

Abstract

The presence of bacterial panicle blight disease caused by Burkholderia glumae have been reported to infect rice plants in Indonesia. There have been no reports of yield losses due to B. glumae infection, but this pathogen is seed-borne so that it can increase the spreading potential to other areas. This study aims to determine the spread of the disease area of bacterial panicle blight in Java and to detect B. glumae based on ITS region of 16-23S rDNA. Methods for this research include surveys and random sampling of rice seeds directly from farmers and the Food Crop and Horticulture Center, Agriculture Office of Yogyakarta. The bacteria were isolated using S-PG selective medium, then DNA extraction was performed and amplified using a pair of primers BGF 5’-ACACGGAACACCTGGGTA-3’ and BGR 5’-TCGCTCTCC CGAAGAGAT-3’. 101 isolates were obtained from 21 seed samples consisting of 11 rice varieties from nine regions in Java that has different morphological characteristics. The eight isolates were detected as B. glumae by using ITS primers, i.e., isolates ChgCM.4, IRP.3, IRP.6b, InSB.1a, InSB.2a, InSB.3a, InSB.5a, and InSB.6a. The eight isolates were isolated from seed samples of Cirebon, Purworejo and Banyuwangi. This study shows that bacterial panicle blight disease has found in several rice varieties and locations in Java, and the ITS primer can be used for early detection of B. glumae in rice seed samples. IntisariPenyakit hawar malai yang disebabkan oleh bakteri patogen Burkholderia glumae mulai banyak dilaporkan menginfeksi tanaman padi di Indonesia. Belum ada laporan mengenai kerugian akibat infeksi B. glumae di Indonesia, namun sifatnya yang tular benih dapat meningkatkan potensi menyebar ke wilayah lain. Penelitian ini bertujuan untuk mengetahui wilayah sebar penyakit hawar malai di Jawa dan mendeteksi secara molekuler berdasarkan urutan basa gen 16-23S rDNA internal transcribed spacer (ITS). Metode yang dilakukan meliputi survei dan pengambilan sampel biji padi secara acak langsung dari petani dan UPT Balai Pengembangan Perbenihan Tanaman Pangan dan Hortikultura, Dinas Pertanian Yogyakarta. Sampel biji padi diisolasi menggunakan media selektif S-PG, kemudian dilakukan ekstraksi DNA, dan diamplifikasi menggunakan primer BGF 5’-ACACGGAACACCTGGGTA-3’ dan BGR 5’-TCGCTCTCC CGAAGAGAT-3’. Hasil survei di lapangan diperoleh 21 sampel biji yang terdiri dari 11 varietas padi dari sembilan wilayah di Jawa. Hasil isolasi diperoleh 101 isolat dan masing-masing isolat tersebut memiliki karakteristik morfologi yang berbeda. Delapan isolat dari total isolat yang diuji terdeteksi B. glumae menggunakan primer ITS, yaitu isolat ChgCM.4, IRP.3, IRP.6b, InSB.1a, InSB.2a, InSB.3a, InSB.5a, dan InSB.6a. Kedelapan isolat tersebut merupakan hasil isolasi sampel biji dari wilayah Cirebon, Purworejo, dan Banyuwangi. Hasil penelitian menunjukkan bahwa penyakit hawar malai bakteri sudah terdeteksi pada varietas padi di beberapa lokasi wilayah Jawa, dan penggunaan primer ITS dapat digunakan untuk deteksi dini B. glumae pada sampel biji padi secara molekuler.