期刊
EUROPEAN JOURNAL OF PLANT PATHOLOGY
卷 156, 期 4, 页码 1207-1212出版社
SPRINGER
DOI: 10.1007/s10658-020-01950-w
关键词
Kiwifruit; Viruses; Multiplex RT-PCR; Detection
资金
- National Natural Science Foundation of China [31772131, 31270290]
- Applied Basic Research Fund of Sichuan Province [2019YJ0135]
- Fundamental Research Funds for the Central Universities [SCU2019D013]
Kiwifruit (Actinidia spp.) is an important horticultural crop in the world. Previous studies showed that a number of viruses were found in kiwifruit around the world. To detect the multiple viruses in kiwifruit quickly and effectively, a multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) method was developed to detect four viruses simultaneously in kiwifruit: Actinidia chlorotic ringspot-associated virus (AcCRaV), Actinidia virus 1(AcV-1), Actinidia virus A (AcVA), Citrus leaf blotch virus (CLBV), respectively. Four pairs of primers were designed according to the coat protein (CP) genes of the aforementioned viruses and then were confirmed by RT-PCR assay. Actin gene (ACT1) was used as an internal control to improve reliability of the multiplex RT-PCR. Determination of detection limit of the multiplex RT-PCR showed that this method could detect four viruses at the dilution of 10(-4) cDNA. Furthermore, the developed multiplex RT-PCR could detect viruses in kiwifruit field samples successfully.
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