Abstract
Monitoring the sanitary status of a large population of vegetative propagation materials is a challenge because it requires expensive and time-consuming testing. Here we report a group testing strategy to identify individual plants carrying viruses and/or a viroid in Pyrus betulifolia seedlings and virus-tested pear mother plants. First, the effect of pool size on RT-PCR detection of apple stem grooving virus (ASGV), apple stem pitting virus (ASPV), apple chlorotic leaf spot virus (ACLSV) and apple scar skin viroid (ASSVd) was evaluated; all of them could be sensitively detected in a bulk (pool) of eight samples. The virological condition of 2770 P. betulifolia seedlings and 110 virus-tested pear mother plants were then investigated by RT-PCR using a grouping strategy with 5 samples. We identified 20, 35 and 3 P. betulifolia seedlings as positive for ASGV, ASPV, and ASSVd, respectively, and 4 pear mother plants were identified as carrying ASGV. Our data indicated that RT-PCR testing on samples pooled before RNA extraction is a reliable and resource-efficient strategy to monitor the sanitary status of large numbers of pear vegetative propagation materials. The results presented here also highlight the possibility of pear seedling rootstocks being a source of viral infection.
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Funding
This work was supported by National Key Research and Development Project of China (2019YFD1001803), the earmarked fund for China Agriculture Research System (CARS-28) and the Construction of Beijing Science and Technology Innovation and Service Capacity in Top Subjects (CEFF-PXM2019_014207_000032).
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FJ and YC contributed to conceptualization; FJ and HL performed experiments; HC, YX, JW, QK contributed to resources; SY analyzed the data; YC contributed to writing original draft preparation; TL contributed to funding acquisition; FJ, HL, HC, YX, JW, QK, SY, TL, YC contributed to Writing review and editing. All authors have read and agreed to the published version of the manuscript.
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Figure S1
Virus (viroid) positive samples identified by group testing.Supplementary file1 (PDF 228 kb)
Table S1
Recombinant plasmids containing the targeted sequence of virus (viroid) used as positive controls in PCR assay.Supplementary file2 (PDF 98 kb)
Table S2
Nucleotide sequences of randomly selected positive PCR products.Supplementary file3 (PDF 147 kb)
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Jiang, F., Li, H., Xie, Y. et al. Identification of individual plants carrying viruses or a viroid in Pyrus betulifolia seedlings and virus-tested pear mother plants by group testing. J Plant Dis Prot 131, 971–976 (2024). https://doi.org/10.1007/s41348-023-00850-5
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DOI: https://doi.org/10.1007/s41348-023-00850-5