PHYCI_587572: An RxLR Effector Gene and New Biomarker in A Recombinase Polymerase Amplification Assay for Rapid Detection of Phytophthora cinnamomi
by
Tingting Dai 1,*,†
, Aohua Wang 1,†, Xiao Yang 2,3, Xiaowei Yu 1, Wen Tian 4, Yue Xu 1 and Tao Hu 1
Tingting Dai 1,*,†, Aohua Wang 1,†, Xiao Yang 2,3, Xiaowei Yu 1, Wen Tian 4, Yue Xu 1 and Tao Hu 1
1
Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China
2
Foreign Disease-Weed Science Research Unit, USDA, Agricultural Research Service (ARS), Fort Detrick, MD 21702, USA
3
ARS Research Participation Program, Oak Ridge Institute for Science and Education, Oak Ridge, TN 37830, USA
4
Compositive Technology Service Center of Jiangyin Customs, Jiangyin 214400, China
*
Author to whom correspondence should be addressed.
†
These authors have contributed equally to this work.
Forests 2020, 11(3), 306; https://doi.org/10.3390/f11030306
Received: 22 January 2020 / Revised: 25 February 2020 / Accepted: 8 March 2020 / Published: 11 March 2020
(This article belongs to the Special Issue Forest Pathology and Entomology)
Phytophthora cinnamomi is a devastating pathogen causing root and crown rot and dieback diseases of nearly 5000 plant species. Accurate and rapid detection of P. cinnamomi plays a fundamental role within the current disease prevention and management programs. In this study, a novel effector gene PHYCI_587572 was found as unique to P. cinnamomi based on a comparative genomic analysis of 12 Phytophthora species. Its avirulence homolog protein 87 (Avh87) is characterized by the Arg-Xaa-Leu-Arg (RxLR) motif. Avh87 suppressed the pro-apoptotic protein BAX- and elicitin protein INF1-mediated cell death of Nicotiana benthamiana. Furthermore, a recombinase polymerase amplification-lateral flow dipstick detection assay targeting this P. cinnamomi-specific biomarker was developed. While successfully detected 19 P. cinnamomi isolates of a global distribution, this assay lacked detection of 37 other oomycete and fungal species, including P. parvispora, a sister taxon of P. cinnamomi. In addition, it detected P. cinnamomi from artificially inoculated leaves of Cedrus deodara. Moreover, the RPA-LFD assay was found to be more sensitive than a conventional PCR assay, by detecting as low as 2 pg of genomic DNA in a 50-µL reaction. It detected P. cinnamomi in 13 infested soil samples, while the detection rate was 46.2% using PCR. Results in this study indicated that PHYCI_587572 is a unique biomarker for detecting P. cinnamomi. Although PHYCI_587572 was identified as an effector gene based on the RxLR motif of Avh87 and the avirulence activity on Nicotiana, its exact genetic background and biological function on the natural hosts of P. cinnamomi warrant further investigations.
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Keywords:
plant destroyers; disease diagnosis; RxLR-dEER; soil-borne pathogen; exclusivity; inclusivity
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MDPI and ACS Style
Dai, T.; Wang, A.; Yang, X.; Yu, X.; Tian, W.; Xu, Y.; Hu, T. PHYCI_587572: An RxLR Effector Gene and New Biomarker in A Recombinase Polymerase Amplification Assay for Rapid Detection of Phytophthora cinnamomi. Forests 2020, 11, 306.
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