Phytophthora cinnamomi causes crown and root wilting in more than 5,000
plant species and represents a significant threat to the health of natural
ecosystems and horticultural crops. The early and accurate detection of P.
cinnamomi is a fundamental step in disease prevention and appropriate
management. In this study, based on public genomic sequence data and
bioinformatic analysis of several Phytophthora, Phytopythium, and Pythium
species, we have identified a new target gene, Pcinn13739; this allowed us to
establish a recombinase polymerase amplification–lateral flow dipstick (RPALFD) assay for the detection of P. cinnamomi. Pcinn13739-RPA-LFD assay was
highly specific to P. cinnamomi. Test results f... More
Phytophthora cinnamomi causes crown and root wilting in more than 5,000
plant species and represents a significant threat to the health of natural
ecosystems and horticultural crops. The early and accurate detection of P.
cinnamomi is a fundamental step in disease prevention and appropriate
management. In this study, based on public genomic sequence data and
bioinformatic analysis of several Phytophthora, Phytopythium, and Pythium
species, we have identified a new target gene, Pcinn13739; this allowed us to
establish a recombinase polymerase amplification–lateral flow dipstick (RPALFD) assay for the detection of P. cinnamomi. Pcinn13739-RPA-LFD assay was
highly specific to P. cinnamomi. Test results for 12 isolates of P. cinnamomi
were positive, but negative for 50 isolates of 25 kinds of Phytophthora species,
13 isolates of 10 kinds of Phytopythium and Pythium species,
32 isolates of 26 kinds of fungi species, and 11 isolates of two kinds of
Bursaphelenchus species. By detecting as little as 10 pg.µl−1 of genomic DNA
from P. cinnamomi in a 50-µl reaction, the RPA-LFD assay was 100 times more
sensitive than conventional PCR assays. By using RPA-LFD assay, P. cinnamomi
was also detected on artificially inoculated fruit from Malus pumila, the leaves
of Rhododendron pulchrum, the roots of sterile Lupinus polyphyllus, and the
artificially inoculated soil. Results in this study indicated that this sensitive,
specific, and rapid RPA-LFD assay has potentially significant applications to
diagnosing P. cinnamomi, especially under time- and resourcelimited conditions.