Identification of Oyster Mushroom Spherical Virus in Pleurotus ostreatus by Deep Sequencing

doi:10.13560/j.cnki.biotech.bull.1985.2014.12.014

Abstract

Abstract: To identify viruses in malformed Pleurotus ostreatus samples collected in Shunyi district in Beijing, deep sequencing of small RNA from total RNA was performed. Consequently, there were 3 075 small interfering RNAs(siRNAs)pairing to the complete sequence of Oyster mushroom isometric virus(OMSV)(NC_004560.1). These siRNAs were located in most regions on genome of OMSV, some of which formed hot-spots. By putting these siRNAs together, three contigs with length more than 500 nucleotides were assembled. According to the sequence of one of these contigs, seq22, a pair of primers was designed to amplify partial coat protein gene via RT-PCR. The results showed that cloned sequence was similar to the reference sequence and to the contig with identities of 91% and 99%, respectively. These results confirmed OMSV in the collected mushroom samples. Meanwhile, the related data acquired from deep sequencing was compared to the predicted siRNAs obtained by four softwares, showing that they were different from each other. The difference was discussed.

Key words: Pleurotus ostreatus, Deep sequencing, Oyster mushroom spherical virus, siRNA, Contig

Wang Shaojie,Wang Nian,Shi Yingchun,Hu Xiaoyan,Zhou Tao. Identification of Oyster Mushroom Spherical Virus in Pleurotus ostreatus by Deep Sequencing[J]. Biotechnology Bulletin, 2014, 0(12): 86-91.

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