Polymorphism in the internal transcribed spacer (ITS) region of the ribosomal DNA among differentFusariumspecies

Abstract

Fusarium species causing wilt diseases in different plants were characterised by comparing nonpathogenic and different pathogenic species using rDNA RFLP analysis. The ITS (internal transcribed spacer) region of 12 isolates belonging to the section Elegans, Laseola, Mortiella, Discolor, Gibbosum, Lateritium and Sporotrichiella were amplified by universal ITS primers (ITS-1 and ITS-4) using polymerase chain reaction (PCR). Amplified products, which ranged from 522 to 565 bp were obtained from all 12 Fusarium isolates. The amplified products were digested with seven restriction enzymes, and restriction fragment length polymorphism (RFLP) patterns were analysed. A dendrogram derived from PCR-RFLP analysis of the rDNA region divided the Fusarium isolates into three major groups. Assessment of molecular variability based on rDNA RFLP clearly indicated that Fusarium species are heterogeneous and most of the forma speciales have close evolutionary relationships.

Molecular Characterization of Fusarium oxysporum Causing Chickpea Wilt by Internal Transcribed Spacer (ITS) Marker

“…These generated two fragments of 220bp and 82bp (Fig 2), 240bp and 52bp (Fig 3), 152bp and150bp (Fig 4) respectively in all isolates except Foc-30, Foc-36 isolates which have not digested by and Foc-25 isolates did not digest by HaeIII. The results obtained from restriction digestion of ITS region were in concordance with earlier studies with different restriction enzymes (Alymanesh et al, 2009;Datta et al, 2011). Two of the restriction enzymes (MspI and AluI) were not digested the ITS region in this study.…”

“…Foc-25 and Foc-48 belonging to Shahartakli, Shelur, Nashik, Satara, Kolhapur respectively and more diverse among the Foc isolates whereas isolate no. Foc 20, Foc 21 and Foc-8 from Ambajogai, Karjat, and Rahata shown minimum genetic distance (0.13) Datta et al (2011). used digested fragments obtained with EcoRI, MspI, PstI, BamHI, XhoI, NstI and HindIII restriction enzymes to determine genetic distances between the pathogenic F. oxysporum isolates of various legumes and clustered them into three main groups Dubey et al (2010).…”

Molecular characterization and vegetative compatibility groups of <i>Fusarium</i> spp. pathogenic to <i>Ilex paraguariensis</i> A. St.-Hil.

“…According to Visentin et al (2009), the sequencing of the ITS region was effective in distinguishing Fusarium verticillioides and Fusarium proliferatum, collected in maize producing regions in northwestern Italy. According to Datta et al (2011) the use of specific primers for the ITS1 and ITS2 regions was effective in distinguishing 12 isolates from Fusarium spp. obtained from different regions of India.…”

TEKNIK PCR ITS-RFLP UNTUK SELEKSI ISOLAT JAMUR PADA PENGUJIAN AGEN PENGENDALI HAYATI PADA SERANGAN GANODERMA TANAMAN Acacia mangium

“…Fragmen ITS banyak digunakan untuk identifikasi maupun karakterisasi jamur karena mudah diamplifikasi, terkonservasi, konsisten, serta memiliki variasi tinggi dalam satu genus, sehingga akan menghasilkan amplikon yang spesifik (Shamim et al, 2017 (Srivastava, Gupta, Lal, & Sinha, 2017) dan sebaliknya akan menghasilkan pola yang sama pada sampel yang memiliki susunan DNA yang serupa (Viaud, Pasquier, & Brygoo, 2000). Penggabungan dari kedua PCR ITS dan RFLP tersebut memungkinkan dihasilkannya penanda yang bersifat spesifik ditingkat spesies, dan memiliki polimorfisme yang tinggi pada isolat jamur tertentu misal Fusarium (Datta, Choudhary, Shamim, & Dhar, 2011) dan Ganoderma (Nusaibah et al, 2011). Penanda ini juga terbukti murah, mudah digunakan dan tidak memerlukan waktu yang lama pada studi keragaman jenis jamur melalui proses isolasi jamur (Diguta et al, 2011), maupun tanpa melalui tahapan isolasi jamur (Viaud et al, 2000).…”