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Siti Subandiyah
Department of Crop Protection, Faculty of Agriculture, Universitas Gadjah Mada Jln. Flora 1, Bulaksumur, Sleman, Yogyakarta 55281
Agriculture, Biological Sciences & Forestry

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Morphological and Molecular Characterization of Rhizoctonia solani Isolates from Two Different Rice Varieties Aisyah Surya Bintang; Arif Wibowo; Achmadi Priyatmojo; Siti Subandiyah
Jurnal Perlindungan Tanaman Indonesia Vol 21, No 2 (2017)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.25469

Abstract

Six isolates of Rhizoctonia solani, i.e. two isolates collected from infected rice plants and four isolates from laboratory collection were studied by using morphological characters and molecular analysis. Un-weighted pair group method with arithmetic mean dendogram constructed based on cluster analysis showed that these isolates were grouped into three clusters at the 0.77 similarity coefficient. Cluster I consisted of BA, BNJ, and NBR isolates with 100% similarity and indicated that those were from AG 1 IA sub group, cluster II consisted of BND, and cluster III consisted of SL1 and SL2. Mycelium was very light brown or whitish with few and moderate sclerotia except SL1 and SL2. Molecular characterization showed that BA, BNJ, and NBR were amplified at 140 bp using Rs1F/Rs2R specific primer for R. solani AG1 IA. All isolates were amplified between 350−400 bp using Rhsp1 primer, meanwhile SL1 and SL2 were not amplified using AG2sp and AG22sp2 primers. Based on Maximum Likelihood tree analysis showed that SL1 and SL2 had high similarity based on ITS sequence data.IntisariEnam isolat Rhizoctonia solani yang berasal dari tanaman padi bergejala dan koleksi laboratorium diuji secara morfologi dan molekuler. Analisis UPGMA dengan koefisien persamaan 0,77 menunjukkan bahwa enam isolat tersebut terbagi atas tiga klaster. Klaster I terdiri atas isolat BA, BNJ, dan NBR dengan kesamaan 100% dan menunjukkan bahwa isolat tersebut berasal dari subgrup AG 1 IA , klaster II yakni isolat BND, dan klaster III terdiri atas isolat SL1 dan SL2. Miselium berwarna putih hingga cokelat muda dengan jumlah sklerotia sedang, kecuali isolat SL1 dan SL2. Uji keragaman secara molekuler menunjukkan bahwa isolat BA, BNJ, dan NBR teramplifikasi pada kisaran 140 bp dengan menggunakan  primer Rs1F/Rs2R yang merupakan primer spesifik dari R. solani AG1 IA. Seluruh isolat teramplifikasi pada kisaran 350−400 bp dengan menggunakan primer Rhsp1, sedangkan isolat SL1 dan SL2 keduanya tidak teramplifikasi oleh primer AG2sp dan AG22sp2. Analisis Maximum Likelihood tree berdasar data sekuen ITS menunjukkan bahwa isolat SL1 dan SL2 memiliki tingkat kesamaan yang tinggi.
Characterization of Pantoea ananatis Isolated from Garlic and Shallot Nanik Nurjanah; Tri Joko; Siti Subandiyah
Jurnal Perlindungan Tanaman Indonesia Vol 21, No 2 (2017)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.27407

Abstract

The new disease on garlic (Allium sativum) and shallot (A. cepa L. aggregatum group) have been found in several production centers of garlic and shallot in Tawangmangu and Temanggung, Central Java. The infected plants showed symptoms of leaf blight accompanied by chlorosis. The objective of this study was to determine the pathogen that causes leaf blight and chlorosis based on the phenotypic characterization and gyrB gene sequences analysis. The research started from the isolation of pathogen, physiological and biochemical test, DNA extraction, and sequence analysis of gyrB using gyrB 01-F and gyrB 02-R primer. The results showed that the isolated bacterial pathogen have a yellow pigment, slimy colonies with regular borders, convex, gram-negative, non-spore, facultative anaerobic, motile, catalase production, indole production, and acid production from D-glucose, D-mannitol, sucrose, and lactose. From the pathogenicity test, it was found that the bacteria produced the typical symptom of leaf blight. Characterization of pathogens based on gyrB gene sequence revealed that the pathogen was placed in the group of Pantoea ananatis. IntisariPenyakit baru pada bawang putih (Allium sativum) dan bawang merah (A. cepa L. aggregatum group) telah ditemukan di beberapa sentra produksi bawang putih dan bawang merah di Tawangmangu dan Temanggung, Jawa Tengah. Tanaman yang terinfeksi menunjukkan gejala hawar daun disertai klorosis. Tujuan penelitian untuk mengetahui karakter patogen berdasarkan fenotipik dan sekuen gen gyrB. Penelitian dimulai dengan isolasi bagian tanaman yang sakit, uji fisiologi dan biokimia, ekstraksi DNA dengan metode CTAB/NaCl dan amplifikasi gen gyrB menggunakan primer gyrB 01-F and gyrB 02-R. Hasil uji menunjukkan koloni berlendir, cembung, pigmen berwarna kuning, gram negative, tidak berspora, aerob fakultatif, motil, produksi katalase, indol, membentuk asam dari D-glukosa, D-monnitol, sukrosa dan laktosa, dan patogenesitas positif. Karakterisasi patogen berdasarkan sekuen gen gyrB, menunjukkan patogen hawar daun berkerabat dekat dengan Pantoea ananatis.
Co-Authors Achmadi Priyatmojo Aisyah Surya Bintang Arif Wibowo Nanik Nurjanah Tri Joko