Reverse Transcription Loop-mediated Isothermal Amplification Assay for Detecting Peach Latent Mosaic Viroid in Peach Pollen

Leehyojeong,; Jeong, Rae‐Dong

doi:10.7235/hort.20200075

Cited by 3 publications

(1 citation statement)

References 28 publications

(30 reference statements)

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“…The detection of other viroids using RT-LAMP was reported to be distinctly rapid where the turbidity was observed within 20-40 min for Pepper chat fruit viroid (PCFVd) (Boubourakas and Kyriakopoulou, 2022;Tangkanchanapas et al, 2018). However, some observations could detect in 50 min for PSTVd (Verma et al, 2020) and 60 min for Peach latent mosaic viroid (PLMVd) (Lee and Jeong, 2020). Hence, the duration depends on the viroid species and their genome sequence (Bhuvitarkorn et al, 2019).…”

Section: P R E S Smentioning

confidence: 99%

Comparison of Real-Time Pcr, Conventional PCR and Rt-Lamp for the Detection of Coconut Cadang-Cadang Viroid Variant in Oil Palm

ROSLAN¹

2022

JOPR

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Real-time quantitative polymerase chain reaction (real-time PCR) was designed for the detection of oil palmCoconut Cadang-cadang Viroid (CCCVd) 246 nt variant. The primers and probe specifically designed for the real-time PCR were optimised with 300 nM of probe against a primer concentration of 400 nM for the detection of oil palm CCCVd variants. Oil palm CCCVd 246 nt variant was successfully detected using realtime PCR in leaf samples collected from 14 symptomatic oil palm from various regions. The sensitivity of real-time PCR was compared with reverse transcription loop-mediated isothermal amplification (RT-LAMP) and conventional PCR. The latter two techniques were reported earlier to be able to detect CCCVd variants in oil palm. Conventional PCR analysis using CCCVd full-length primers detected the presence of CCCVd 246 nt variant in 10 while RT-LAMP detected only seven positive samples from the total 14 field samples.The real-time PCR was highly sensitive and reliable compared to the two detection techniques. The primers designed for the real-time PCR were specific as only the oil palm CCCVd variant plasmid was detected with high fluorescence and lowest quantification cycle (Cq value) compared to the other tested viroids (ASSVd, CTiVd, ELVd, HLVd and PLMVd). The present study has proven the reliability of the technique, thus, highly recommending the next step of developing the high-throughput diagnostic procedure for the eventual use in epidemiological monitoring programmes.

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Section: P R E S Smentioning

confidence: 99%

Comparison of Real-Time Pcr, Conventional PCR and Rt-Lamp for the Detection of Coconut Cadang-Cadang Viroid Variant in Oil Palm

ROSLAN¹

2022

JOPR

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Advancements in the loop-mediated isothermal amplification technique for the rapid detection of plant viruses in various crops

Fu,

Sha,

et al. 2024

Physiological and Molecular Plant Pathology

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Viromes of Hungarian Peach Trees Identified by High-Throughput Sequencing of Small RNAs

Baráth

Jaksa-Czotter

Varga

et al. 2022

Plants

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Peach trees can be infected with viruses and viroids. As we do not have efficient plant protection methods against these pathogens, the prevention of infection is crucial. Fruit trees are maintained by vegetative propagation. Planting material such as certified mother trees and rootstocks should be free from viruses and viroids, and this status has to be regularly checked to prevent infections. We surveyed certified peach trees for the presence of viruses and viroids using small RNA high-throughput sequencing (HTS), an unbiased virus diagnostic method. The results of the bioinformatic analysis of HTS were validated by other molecular methods including RT-PCR, Northern blot hybridization and loop-mediated isothermal amplification (LAMP). We found the presence of plum pox virus and peach latent mosaic viroid (PLMVd) in the vector-free isolator houses, whose presence should be regularly tested. Moreover, we detected frequent infection with recently described viruses such as nectarine stem pitting-associated virus and peach-associated luteovirus (PaLV). During the survey, PLMVd and PaLV were detected for the first time in Hungary. The analysis of the presenting virus variants and possible sources of infection suggests that the source of the viral infection could be the infected propagating material. Our study emphasizes the importance of using sensitive and trustworthy diagnostic techniques to be able to detect viral infections and successfully prevent their spread by propagation material.

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Reverse Transcription Loop-mediated Isothermal Amplification Assay for Detecting Peach Latent Mosaic Viroid in Peach Pollen

Cited by 3 publications

References 28 publications

Comparison of Real-Time Pcr, Conventional PCR and Rt-Lamp for the Detection of Coconut Cadang-Cadang Viroid Variant in Oil Palm

Comparison of Real-Time Pcr, Conventional PCR and Rt-Lamp for the Detection of Coconut Cadang-Cadang Viroid Variant in Oil Palm

Advancements in the loop-mediated isothermal amplification technique for the rapid detection of plant viruses in various crops

Viromes of Hungarian Peach Trees Identified by High-Throughput Sequencing of Small RNAs

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