大蒜病毒多重RT-PCR检测体系的建立与应用

doi:10.12190/j.issn.2096-1197.2023.01.03

Abstract

Abstract: 【Objective】To establish a multiplex RT-PCR system for simultaneous detection and identification of 5 viruses/virus groups in garlic field plants and tissue cultured plantlets.【Methods】Based on relevant literature and the reference virus sequences registered in GenBank,5 sets of specific primers were designed for Shallot latent virus（ShLV）,Garlic common latent virus（GarCLV）,Leek yellow stripe virus（LYSV）,Onion yellow dwarf virus（OYDV）,and allexiviruses,with amplicon size of 592,431,338,265,and 190 bp,respectively. Using the established multiplex RT-PCR method,virus detection and analysis were conducted on 24 garlic tissue cultured plantlets and 48 field samples.【Results】The specificity of the detection system was verified by single/multiplex PCR and sequence alignment. By adjusting the annealing temperature and primer ratio,the optimal amplification effect was achieved at 56.6 ℃ annealing temperature and the primer volume ratio of ShLV,GarCLV,LYSV,OYDV,and allexiviruses at 10∶8∶3∶3∶16. All 5 viruses/virus groups had a minimum detection concentration of 1.0×10² copies/μL. The virus detection results of garlic tissue cultured plantlets were rich in diversity. The field samples were infected with at least 2 viruses/virus groups,and the viruses/virus groups could be clearly distinguished.【Conclusion】The developed multiplex RT-PCR system quickly and effectively detected and differentiated the infection status of the five viruses/virus groups in garlic,including ShLV,GarCLV,LYSV,OYDV,and allexiviruses.

Key words: Garlic, Virus, Multiplex RT-PCR, Detection system

CLC Number:

S436.33

ZHAO Yongqiang, FAN Jide, LU Xinjuan, LIU Canyu, ZHANG Biwei, YANG Qingqing, GE Jie, SHI Xinmin, YANG Feng. Development and application of a multiplex RT-PCR detection system for garlic viruses[J].Journal of Northern Agriculture, 2023, 51(1): 22-30.

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https://journal30.magtechjournal.com/bfnyxb/EN/Y2023/V51/I1/22

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Development and application of a multiplex RT-PCR detection system for garlic viruses

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