| Detection of GLV, OYDV and LYSV in potato onion （Allium cepa L., Aggregatum group） by RT-PCR |
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| 作者姓名： | XU Qijiang CHEN Dian TONG Youli and LI Yuhua* Research Institute of Flower Biotechnology Northeast Forestry University Harbin China College of Horticulture Northeast Agricultural University Harbin China |
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| 作者单位： | XU Qijiang1，CHEN Dian2，TONG Youli1，and LI Yuhua1* 1 Research Institute of Flower Biotechnology，Northeast Forestry University，Harbin 150040，China 2 College of Horticulture，Northeast Agricultural University，Harbin 150030，China |
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| 基金项目： | Supported by Department Education of Heilongjiang Province of China （10531146） |
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| 摘 要： | Three pairs of primers were designed and synthesized from nucleotide sequences of garlic latent virus (GLV), onion yellow dwarf virus (OYDV), and leek yellow stripe virus (LYSV) by using PCR primer design software. The expected fragments about 170 bp, 287 bp, and 191 bp were amplified by RT-PCR for GLV, OYDV, and LYSV, respectively in disease-infected plants of potato onion (Allium cepa L., Aggregatum group), but such fragments were not obtained from healthy-looking plants and virus-free seedlings of shoot-tips. The amplified products of GLV, OYDV and LYSV were cloned into pGEM-T vectors, and transformed into Escherichia coli. JM109. The recombinant plasmids were obtained and sequenced. The nucleotide sequences were compared with corresponding viral nucleotide sequences reported in GenBank by performing a NCBI BLAST. The analysis showed that their homology attained 75% to 90%,89.5% to 96.1%,and 91.6% to 96.3% in GLV, OYDV, and LYSV, respectively. The total RNA of 6.34 μg·μL~(-1) from infected plants was diluted to a series of 10~(-1) to 10~(-5) and the detection sensitivity of RT-PCR was 10~(-4) (about 4 ng). Thus, a method of identification and detection by RT-PCR of GLV, OYDV, and SLYV was established.
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| 关 键 词： | 洋葱病毒病 大蒜潜隐病毒 洋葱黄萎病毒 韭葱黄条斑纹病毒 RT-PCR 检测 |
| 文章编号： | 1006-8104（2007）-01-0009-05 |
| 收稿时间： | 2006-06-26 |
Detection of GLV, OYDV and LYSV in potato onion (Allium cepa L.,Aggregatum group) by RT-PCR |
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| XU Qijiang,CHEN Dian,TONG Youli,and LI Yuhua* Research Institute of Flower Biotechnology,Northeast Forestry University,Harbin ,China College of Horticulture,Northeast Agricultural University,Harbin ,China.Detection of GLV, OYDV and LYSV in potato onion (Allium cepa L.,Aggregatum group) by RT-PCR[J].Journal of Northeast Agricultural University,2007,14(1):9-13. |
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| Authors: | XU Qijiang CHEN Dian TONG Youli LI Yuhua |
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| Abstract: | Three pairs of primers were designed and synthesized from nucleotide sequences of garlic latent virus (GLV), onion yellow dwarf virus (OYDV), and leek yellow stripe virus (LYSV) by using PCR primer design software. The expected fragments about 170 bp,287 bp, and 191 bp were amplified by RT-PCR for GLV, OYDV, and LYSV, respectively in disease-infected plants of potato onion(Allium cepa L., Aggregatum group), but such fragments were not obtained from healthy-looking plants and virus-free seedlings of shoot-tips. The amplified products ofGLV, OYDV and LYSV were cloned into pGEM-T vectors, and transformed into Escherichia coli.JM109. The recombinant plasmids were obtained and sequenced. The nucleotide sequences were compared with corresponding viral nucleotide sequences reported in GenBank by performing a NCBI BLAST. The analysis showed that their homology attained 75% toplants was diluted to a series of 10-1 to 10-5 and the detection sensitivity of RT-PCR was 10-4 (about 4 ng). Thus, a method of identification and detection by RT-PCR of GLV, OYDV, and SLYV was established. |
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| Keywords: | garlic latent virus (GLV) onion yellow virus (OYDV) leek yellow stripe virus (LYSV) RT-PCR |
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