Tissue Blotting and RT-PCR Detection of Soybean Mosaic Virus(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:

2010年06期

Page:

915-919

Research Field:

Publishing date:

Info

Title:

Tissue Blotting and RT-PCR Detection of Soybean Mosaic Virus

Author(s):

ZHAN Yong¹; 2;  YU De-yue¹

1.National Key Laboratory for Crop Genetics and Germplasm Enhancement, National Center for Soybean Improvement, Soybean Research Institute, Nanjing Agricultural University, Nanjing 210095, Jiangsu;

2. Crop Research Institute, Xinjiang Academy of Agriculture and Reclamation Science, Shihezi 832000, Xinjiang, China

Keywords:

Soybean mosaic virus; Tissue blotting; RT-PCR; Detection

PACS:

S565.1

DOI:

10.11861/j.issn.1000-9841.2010.06.0915

Abstract:

With improved tissue blotting approach, 69 SMV samples from the Yellow and Huaihe River areas were detected and the results showed that 42 samples presented a very clear blue-violet-positive reaction, 26 samples could not made conclusion for weaker color and one sample was negative. Further analysis by 600 dpi scan and enlarging the picture for 200 times, it was observed that 25 from 26 samples suffering from weak color showed positive results, and the negative standard sample were still unable to confirm the color, which makes the detection rate increased by 37%. RT-PCR verification were used in random selected eight samples and the results showed that both purified and unpurified RNA templates could obtain distinct bands, however, only the purified PCR products could be amplified repeatedly with the expected secondary bands, and the double-digestion could also produce three expected fragments, 245 bp, 255 bp and 307 bp. The results above indicated that the improved tissue blotting method could detect soybean SMV efficiently and bulkily, RT-PCR detection could be used in more precise testing.

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