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| Establishment of Duplex RT-PCR Detection System for PlAMV and LMoV in Lily (Lilium spp.) |
| WANG Li-Hua1, YANG Xiu-Mei1,*, HE Xiao-Qin2, ZHANG Yi-Ping1, DUAN Qing1, YANG Shu-Qi1, SU Yan1,* |
1 Flower Research Institute/Yunnan Key Laboratory for Flower Breeding/National Engineering Research Center for Ornamental Horticulture,Yunnan Academy of Agriculture Science, Kunming 650205, China;
2 Qingyun subdistrict office Panlong District Government, Kunming 650051, China |
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Abstract The plantago asiatica mosaic virus (PlAMV) and the Lily mottle virus (LMoV) are currently the two most serious viral pathogens affecting the lily (Lilium spp.) industry in China. The aim of this study is to develop a rapid and efficient detection system for these 2 viruses. Firstly, total RNA was extracted from diseased lily plants, and cDNA was obtained through reverse transcription. Subsequently, a dual-synchronous detection system was optimised using cDNA as a template, and the specificity and sensitivity of the detection system were evaluated. The results demonstrated that the established dual-synchronous detection system amplified specific segments of 910 and 521 bp for PlAMV and LMoV, respectively, with no amplification for other common viruses, indicating strong detection specificity. The lowest detection limits for PlAMV and LMoV were 967.5 fg and 59.0 pg, respectively, which were 50 times and 5 times lower than the lowest detection limits of single-gene RT-PCR detection (PlAMV 19.35 fg, LMoV 11.8 pg). Sequence analysis of the amplified products with sequencing segments from 19 PlAMV isolates and 21 LMoV isolates from different sources revealed similarities of 86.71% to 99.40% for PlAMV and 94.75% to 99.49% for LMoV. The dual-synchronous detection system was employed to examine 19 field samples, and the results indicated that 3 samples were positive for both PlAMV and LMoV, 2 samples were positive for PlAMV, 4 samples were positive for LMoV, and the remaining 10 samples were not detected, which was consistent with the results of gene chip detection. In conclusion, the detection system this study established is capable of detecting both PlAMV and LMoV viruses simultaneously, with reliable results and broad application prospects.
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Received: 08 February 2024
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Corresponding Authors:
* 175399925@qq.com; 1119707040@qq.com
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