Early Detection of Both Pyrenophora teres f. teres and f. maculata in Asymptomatic Barley Leaves Using Digital Droplet PCR (ddPCR)

被引:0
作者
Bouhouch, Yassine [1 ,2 ]
Aggad, Dina [2 ]
Richet, Nicolas [1 ]
Rehman, Sajid [3 ]
Al-Jaboobi, Muamar [3 ]
Kehel, Zakaria [3 ]
Esmaeel, Qassim [1 ]
Hafidi, Majida [4 ]
Jacquard, Cedric [1 ]
Sanchez, Lisa [1 ]
机构
[1] Univ Reims, INRAE, RIBP, USC 1488, BP 1039, Reims, France
[2] Univ Reims, Plateformes Technol URCATech, Plateau MOBICYTE, BP 1039, Reims, France
[3] Int Ctr Agr Res Dry Areas, Biodivers & Crop Improvement Program, BP 6202, Rabat, Morocco
[4] Univ Moulay Ismail, Fac Sci, Lab Biotechnol Vegetale & Biol Mol, BP 11201, Zitoune, Meknes, Morocco
关键词
barley; Pyrenophora teres; k-mer analysis; qPCR; ddPCR; early detection; POLYMERASE-CHAIN-REACTION; REAL-TIME PCR; NET BLOTCH; SPOT FORM; GENETIC-STRUCTURE; CAUSAL AGENT; PATHOGENS; MARKERS; QUANTIFICATION; POPULATIONS;
D O I
10.3390/ijms252211980
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Efficient early pathogen detection, before symptom apparition, is crucial for optimizing disease management. In barley, the fungal pathogen Pyrenophora teres is the causative agent of net blotch disease, which exists in two forms: P. teres f. sp. teres (Ptt), causing net-form of net blotch (NTNB), and P. teres f. sp. maculata (Ptm), responsible for spot-form of net blotch (STNB). In this study, we developed primers and a TaqMan probe to detect both Ptt and Ptm. A comprehensive k-mer based analysis was performed across a collection of P. teres genomes to identify the conserved regions that had potential as universal genetic markers. These regions were then analyzed for their prevalence and copy number across diverse Moroccan P. teres strains, using both a k-mer analysis for sequence identification and a phylogenetic assessment to establish genetic relatedness. The designed primer-probe set was successfully validated through qPCR, and early disease detection, prior to symptom development, was achieved using ddPCR. The k-mer analysis performed across the available P. teres genomes suggests the potential for these sequences to serve as universal markers for P. teres, transcending environmental variations.
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页数:16
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