AUTHOR=Chen Zhenpeng , Jiao Binbin , Zhou Jing , He Haibin , Dai Tingting 

TITLE=Rapid detection of Phytophthora cinnamomi based on a new target gene Pcinn13739

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2022.923700

DOI=10.3389/fcimb.2022.923700

ISSN=2235-2988

ABSTRACT=<p><italic>Phytophthora cinnamomi</italic> 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 <italic>P. cinnamomi</italic> is a fundamental step in disease prevention and appropriate management. In this study, based on public genomic sequence data and bioinformatic analysis of several <italic>Phytophthora</italic>, <italic>Phytopythium</italic>, and <italic>Pythium</italic> species, we have identified a new target gene, Pcinn13739; this allowed us to establish a recombinase polymerase amplification–lateral flow dipstick (RPA-LFD) assay for the detection of <italic>P. cinnamomi</italic>. <italic>Pcinn13739</italic>-RPA-LFD assay was highly specific to <italic>P. cinnamomi</italic>. Test results for 12 isolates of <italic>P. cinnamomi</italic> were positive, but negative for 50 isolates of 25 kinds of <italic>Phytophthora</italic> species, 13 isolates of 10 kinds of <italic>Phytopythium</italic> and <italic>Pythium</italic> species, 32 isolates of 26 kinds of fungi species, and 11 isolates of two kinds of <italic>Bursaphelenchus</italic> species. By detecting as little as 10 pg.µl<sup>−1</sup> of genomic DNA from <italic>P. cinnamomi</italic> in a 50-µl reaction, the RPA-LFD assay was 100 times more sensitive than conventional PCR assays. By using RPA-LFD assay, <italic>P. cinnamomi</italic> was also detected on artificially inoculated fruit from <italic>Malus pumila</italic>, the leaves of <italic>Rhododendron pulchrum</italic>, the roots of sterile <italic>Lupinus polyphyllus</italic>, 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 <italic>P. cinnamomi</italic>, especially under time- and resource-limited conditions.</p>