Development and Evaluation of a Real-time Fluorescent Quantitative PCR Assay for Detection of Sweet Potato Latent Virus-Lotus in Lotus Plants

doi:10.16420/j.issn.0513-353x.2019-0859

ACTA HORTICULTURAE SINICA ›› 2020, Vol. 47 ›› Issue (7): 1412-1420.doi: 10.16420/j.issn.0513-353x.2019-0859

• New Technologies and New Methods • Previous Articles Next Articles

Development and Evaluation of a Real-time Fluorescent Quantitative PCR Assay for Detection of Sweet Potato Latent Virus-Lotus in Lotus Plants

HE Zhen，DONG Tingting，WU Weiwen，CHEN Wen，and LI Liangjun*

School of Horticulture and Plant Protection，Yangzhou University，Yangzhou，Jiangsu 225009，China

Online:2020-07-25 Published:2020-07-25

Abstract

Abstract: During the survey of lotus viruses disease，sweet potato latent virus-lotus was found to occur at high frequencies in Jiangsu province, China. And there is a significant sequence difference between SPLV-lotus and sweet potato isolates. In order to detect SPLV more accurately，a specific RT-qPCR assay based on SYBR GreenⅠfluorescent dye was established. A specific primer pair QSPLV- lotus3-F/QSPLV-lotus3-R was designed based on the CP gene of SPLV-lotus，and a standard curve was constructed under the optimized primer concentration and annealing temperature. The results showed that the RT-qPCR assay had higher specificity and was 100 times more sensitive for SPLV detection than conventional RT-PCR. This method can be widely used in batch inspection of field samples and be suitable for the detection and identification of virus-free lotus root seedlings.

Key words: lotus, sweet potato latent virus-lotus, RT-qPCR

CLC Number:

S 645.1

HE Zhen，DONG Tingting，WU Weiwen，CHEN Wen，and LI Liangjun*. Development and Evaluation of a Real-time Fluorescent Quantitative PCR Assay for Detection of Sweet Potato Latent Virus-Lotus in Lotus Plants[J]. ACTA HORTICULTURAE SINICA, 2020, 47(7): 1412-1420.

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Development and Evaluation of a Real-time Fluorescent Quantitative PCR Assay for Detection of Sweet Potato Latent Virus-Lotus in Lotus Plants

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