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    FEMS Microbiology Letters发表大豆疫霉LAMP检测技术

      

    题目:Development of a Loop-Mediated Isothermal Amplification assay for detection of phytophthora sojae

    作者:Ting-ting Dai, Chen-Chen Lu, Suo-Meng Dong, Xiao-Bo Zheng ,Yuan-Chao Wang*, Department of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, China

    研究摘要: Phytophthora sojae is a devastating pathogen that causes soybean Phytophthora root rot. This paper reports on the development of a loop-mediated isothermal amplification (LAMP) assay targeting A3aPro gene for visual detection of P. sojae. The A3aPro-LAMP assay efficiently amplified the target gene within 60 min at 64°C and was evaluated for specificity and sensitivity. The specificity of the assay was evaluated against isolates of P. sojae, Phytophthora spp., Pythium spp., and true fungi. Magnesium pyrophosphate resulting from the LAMP of P. sojae could be detected by monitoring Real-time measurement of turbidity; Meanwhile, the DNA products of P. sojae were visualized as a ladder-like banding pattern on 2% gel electrophoresis and a positive color (sky blue) was only observed in the presence of P. sojae by addition of hydroxynaphthol blue prior to the amplification, whereas none of other isolates showed a color change. The detection limit of A3apro-specific LAMP assay was 10 pg µl-1 of genomic DNA per reaction. The LAMP assay was also successfully applied for the detection of P. sojae from diseased soybean tissues and residues. These results suggest that the A3apro-LAMP assay reported here shows great potential for the visual detection of P. sojae in plants and production fields.


    发布时间: 2012-07-05 13:15 浏览次数:   【 打 印 】【 关 闭
     
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