Use of LAMP Detection to Identify Potential Contamination Sources of Plant-Pathogenic Pythium Species in Hydroponic Culture Systems of Tomato and Eustoma

Abstract

Hydroponic culture systems are subject to high risks of diseases caused by zoosporic plant pathogens. Control is generally difficult because of the rapid spread of zoospores in the nutrient solutions. In Japan, tomato and eustoma, which are cultivated using the D-tray and nutrient film techniques, respectively, are susceptible to diseases caused by Pythium aphanidermatum and P. irregulare. We used loop-mediated isothermal amplification to identify potential contamination sources of these two pathogens by monitoring their presence in the water supply wells, seedling terraces, nutrient solutions, diseased plants, and ground soils of a tomato greenhouse complex and a eustoma greenhouse complex. The results indicated that the pathogens may enter the culture systems from the soils around the greenhouses. Entry most likely occurs when seedlings are moved from the seedling terraces to the greenhouses, and sterilization of the hydroponic systems may not be sufficient. Therefore, monitoring pathogens in the culture systems and ground soils is very important for the management and prevention of these diseases.

Next‐generation methods for early disease detection in crops

“…Like PCR, LAMP can detect a single target in a complex matrix, or multiple targets from it. In addition, VLAMP is sufficiently sensitive to detect quarantine pathogens from small amounts of biological traces, such as frass samples 90 or DNA extracted from soil and water samples to monitor soil‐borne pathogens in a greenhouse 91 . More recently, VLAMP detected plant pathogens in pre‐symptomatic and infected plant tissue 92 .…”

“…In addition, VLAMP is sufficiently sensitive to detect quarantine pathogens from small amounts of biological traces, such as frass samples 90 or DNA extracted from soil and water samples to monitor soil-borne pathogens in a greenhouse. 91 More recently, VLAMP detected plant pathogens in pre-symptomatic and infected plant tissue. 92 In reverse transcription LAMP (RT-LAMP), reverse transcriptase step is added to the LAMP mixture to convert RNA samples into cDNA for amplification.…”

Biocontrol potential of endophytic Pseudomonas strain IALR1619 against two Pythium species in cucumber and hydroponic lettuce

“…It is less susceptible to diseases than hydroponic spinach and has a shorter seedling to harvest window compared to fruit bearing crops like tomatoes and cucumber [ 12 ]. However, root rot caused by various Pythium species pose a threat in hydroponic systems since pathogen propagules can easily spread with circulating nutrient solutions and infect plants [ 9 , 10 , 12 , 13 ]. Several Pythium species have been reported to cause root rot and yield reduction in hydroponic lettuce.…”

Preliminary optimisation of simplified sample preparation method to permit direct detection of SARS-CoV-2 within saliva samples using reverse- transcription loop-mediated isothermal amplification (RT-LAMP)

“…Reverse-Transcription Loop-mediated isothermal AMPlification (RT-LAMP) is a highly sensitive reverse-transcription, autocycling, isothermal, strand displacement nucleic acid amplification technology 8 which is more resistant to inhibitors than rRT-PCR), enabling simplification and even removal of the extraction procedure 9–11 . LAMP technologies have been applied for the detection of a wide range of pathogens 12–14 including positive-sense RNA viruses 10 and has been used extensively in the veterinary 15,16 and plant industry 17–19 and more recently as a human diagnostic 12,20,21 . At the height of the SARS-CoV-2 epidemic in the UK in early 2020, Hampshire Hospitals NHS Foundation Trust (HHFT) validated a novel RT-LAMP assay for the detection of SARS-CoV-2 RNA within nasopharyngeal and oropharyngeal swabs either directly from swab, or following RNA extraction 22 .…”