Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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A survey of viruses and viroids in astringent persimmon (Diospyros kaki Thunb.) and the development of a one-step multiplex reverse transcription-polymerase chain reaction assay for the identification of pathogens

  • Kwon, Boram (School of Applied Biosciences, Kyungpook National University) ;
  • Lee, Hong-Kyu (School of Applied Biosciences, Kyungpook National University) ;
  • Yang, Hee-Ji (School of Applied Biosciences, Kyungpook National University) ;
  • Kim, So-Yeon (School of Applied Biosciences, Kyungpook National University) ;
  • Lee, Da-Som (School of Applied Biosciences, Kyungpook National University) ;
  • An, ChanHoon (Forest Microbiology Division, National Institute of Forest Science) ;
  • Kim, Tae-Dong (Forest Tree Improvement and Biotechnology Division, National Institute of Forest Science) ;
  • Park, Chung Youl (Division of Wild Plant Seeds Research, Baekdudaegan National Arboretum) ;
  • Lee, Su-Heon (School of Applied Biosciences, Kyungpook National University)
  • Received : 2022.07.11
  • Accepted : 2022.08.05
  • Published : 2022.09.30
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Abstract

Astringent persimmon (Diospyros kaki Thunb.) is an important fruit crop in Korea; it possesses significant medicinal potential. However, knowledge regarding the pathogens affecting this crop, particularly, viruses and viroids, is limited. In the present study, reverse transcription-polymerase chain reaction (RT-PCR) and high-throughput transcriptome sequencing (HTS) were used to investigate the viruses and viroids infecting astringent persimmons cultivated in Korea. A one-step multiplex RT-PCR (mRT-PCR) method for the simultaneous detection of the pathogens was developed by designing species-specific primers and selecting the primer pairs via combination and detection limit testing. Seven of the sixteen cultivars tested were found to be infection-free. The RT-PCR and HTS analyses identified two viruses and one viroid in the infected samples (n = 51/100 samples collected from 16 cultivars). The incidence of single infections (n = 39/51) was higher than that of mixed infections (n = 12/51); the infection rate of the Persimmon cryptic virus was the highest (n = 31/39). Comparison of the monoplex and mRT-PCR results using randomly selected samples confirmed the efficiency of mRT-PCR for the identification of pathogens. Collectively, the present study provides useful resources for developing disease-free seedlings; further, the developed mRT-PCR method can be extended to investigate pathogens in other woody plants.

Keywords

Acknowledgement

This research was funded by the National Institute of Forest Science (grant number FG0700-2018-02-2021). We would like to thank Editage (https://www.editage.co.kr) for English language editing.

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