Amplifying and Sequencing Analysis the Internal Transcribed Spacer (ITS) Regions of Olpidium Viciae Kusano’s Ribosomal DNA in Broad Bean

Abstract

Plasmodiophora fire of broad bean is responsible for Olpidium Viciae Kusano, which is a kind of Fungi subdivided into bacteria flagellum amon. We have developed a polymerase chain reaction based method for the rapid identification internal transcribed spacer (ITS) regions of productionally significant fungi Olpidium Viciae Kusano from areas of 2500~3000 metres above sea level. Sequences of the nuclear internal transcribed spacer (ITS) regions ITS1 and ITS4 have been used widely in molecular characteristic studies because of their relatively high variability and facility of amplification. A universal quickly SDS micro-DNA extraction method was used combining a RNaseA pretreatment step to remove PCR interferential RNA. Target sequences in ITS regions genomic were amplified by PCR and sequenced. Using Hanpanchun lesion and healthy bean leaves as template and ITS1, ITS4 as primer to amplify ITS region, the results revealed ITS gene of broad bean genome could be amplified with size of 750bp from healthy leaves, it could be amplified two fragments of 750bp and 500bp from the DNA template extracted from Hanpanchun lesion tissue. The ITS sequence of Olpidium Viciae Kusano is 99% homoeology with Cercospora (grey speck) pathogen. This may lay the foundation for research about classification and analyze evolutionary relationships of Olpidium Viciae Kusano.

Potential of marine sponge-derived fungi in the aquaculture system

“…Table 3 shows the result of homology analysis of the isolate sequence of SL 3 SP 3.3 which has a sequence length of 638 bp and has a 99.69% similarity with the Trichoderma reesei RHa strain under the accession number KM246746.1. Primers covered the sequence length of the fungal isolate ITS 1 and ITS 4 with amplification ranged 750bp and 500bp (Yan et al 2011). ITS primer is a primer that matched 99% of ascomycete and basidiomycete taxa (species, subspecies, or varieties) based on public sequence databases named in silico analysis (Toju et al 2012).…”

Faba Bean Gall (Olpidium viciae K.) as a Priority Biosecurity Threat for Producing Faba Bean in Ethiopia: Current Status and Future Perspectives

“…Yan et al [110] did rapid identification of internal transcribed spacer (ITS) regions of O. viciae by developing a polymerase chain reaction-based method.…”

Identifying phytopathogenic fungi in Al-Baha province, Saudi Arabia through their molecular and morphological features: An overview

“…Within ITS regions the most extensively favoured primers used for fungal molecular identification are the ITS1 and ITS4 primers due to their large coverage between the small subunit 18S and the large subunit 28S, which showed a high level of variation between different species ( Fig. 1 ) ( Yan et al, 2011 ). By 2012, the gene banks held more than 175,000 lengths of ITS sequences, representing over 15,000 fungal species ( Schoch et al, 2012 ).…”

“…1 Illustrating the position of the internal transcribed spacer region (ITS), which is located between the small and the large ribosomal subunits (18S and 28S rRNA). The arrows indicate the coverage area for the forward primer, ITS1, and the reverse primer, ITS4 ( Yan et al, 2011 ). …”