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A Duplex PCR Assay for Rapid Detection of Phytophthora nicotianae and Thielaviopsis basicola.


ABSTRACT: A duplex PCR method was developed for simultaneous detection and identification of tobacco root rot pathogens Phytophthora nicotianae and Thielaviopsis basicola. The specific primers for P. nicotianae were developed based on its internal transcribed spacer (ITS) regions of ribosomal gene, ras gene and hgd gene, while the specific primers for T. basicola were designed based on its ITS regions and ?-tubulin gene. The specificity of the primers was determined using isolates of P. nicotianae, T. basicola and control samples. The results showed that the target pathogens could be detected from diseased tobacco plants by a combination of the specific primers. The sensitivity limitation was 100 fg/?l of pure genomic DNA of the pathogens. This new assay can be applied to screen out target pathogens rapidly and reliably in one PCR and will be an important tool for the identification and precise early prediction of these two destructive diseases of tobacco.

SUBMITTER: Liu N 

PROVIDER: S-EPMC6464196 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature

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A Duplex PCR Assay for Rapid Detection of <i>Phytophthora nicotianae</i> and <i>Thielaviopsis basicola</i>.

Liu Na N   Jiang Shijun S   Feng Songli S   Shang Wenyan W   Xing Guozhen G   Qiu Rui R   Li Chengjun C   Li Shujun S   Zheng Wenming W  

The plant pathology journal 20190401 2


A duplex PCR method was developed for simultaneous detection and identification of tobacco root rot pathogens <i>Phytophthora nicotianae</i> and <i>Thielaviopsis basicola</i>. The specific primers for <i>P. nicotianae</i> were developed based on its internal transcribed spacer (ITS) regions of ribosomal gene, <i>ras</i> gene and <i>hgd</i> gene, while the specific primers for <i>T. basicola</i> were designed based on its ITS regions and <i>β-</i>tubulin gene. The specificity of the primers was d  ...[more]

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