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Establishment and Application of a Multiplex PCR Assay for the Rapid Detection of Rhizoctonia solani Anastomosis Group (AG)-3PT, the Pathogen Causing Potato Black Scurf and Stem Canker

Iradukunda, Linda and Wang, Yan-Ping and Nkurikiyimfura, Oswald and Wang, Tian and Yang, Li-Na and Zhan, Jiasui (2022). Establishment and Application of a Multiplex PCR Assay for the Rapid Detection of Rhizoctonia solani Anastomosis Group (AG)-3PT, the Pathogen Causing Potato Black Scurf and Stem Canker. Pathogens. 11 :6 , 627
[Research article]

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Abstract

Rhizoctonia solani anastomosis group 3 (AG-3) is the main causative agent of the soil-borne disease known as potato black scurf, which poses a huge threat to potato production. Rapid and accurate identification of R. solani AG-3 isolates in soil and potato seed tubers prior to planting is essential for good production. In this study, a multiplex PCR assay was established for the detection of R. solani AG-3. Two pairs of target-specific primers were designed from sequences for endopolygalacturonase and pyridoxine biosynthesis genes downloaded from GenBank. The main factors influencing PCR amplification, such as annealing temperature and primer concentration, were optimized. Results show that the proposed multiplex PCR assay is highly sensitive and specific for the target genes in the pathogen even when the DNA concentration is reduced to 20 fg/mu L. The resulting calibration plot shows a linear relationship between electrophoretic band peaks and genomic DNA concentration (R-2 = 0.98). The primer specificity was confirmed by applying them to other R. solani AG groups and plant pathogen species on which no amplicons were produced. Using the primers, we successfully detected small amounts of R. solani AG-3 present in soil and potato tuber samples. Taken together, the detection assay developed in this study has high sensitivity, strong specificity, and accuracy and can be used to detect and identify soil and potato seed tubers infected with Rhizoctonia solani AG-3.

Authors/Creators:Iradukunda, Linda and Wang, Yan-Ping and Nkurikiyimfura, Oswald and Wang, Tian and Yang, Li-Na and Zhan, Jiasui
Title:Establishment and Application of a Multiplex PCR Assay for the Rapid Detection of Rhizoctonia solani Anastomosis Group (AG)-3PT, the Pathogen Causing Potato Black Scurf and Stem Canker
Series Name/Journal:Pathogens
Year of publishing :2022
Volume:11
Number:6
Article number:627
Number of Pages:11
Publisher:MDPI
Language:English
Publication Type:Research article
Article category:Scientific peer reviewed
Version:Published version
Copyright:Creative Commons: Attribution 4.0
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Microbiology (Microbiology in the medical area to be 30109)
Keywords:potato, Rhizoctonia solani AG-3, multiplex PCR, molecular detection
URN:NBN:urn:nbn:se:slu:epsilon-p-118287
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-118287
Additional ID:
Type of IDID
DOI10.3390/pathogens11060627
Web of Science (WoS)000816692100001
ID Code:28697
Faculty:NJ - Fakulteten för naturresurser och jordbruksvetenskap
Department:(NL, NJ) > Dept. of Forest Mycology and Plant Pathology
(S) > Dept. of Forest Mycology and Plant Pathology
Deposited By: SLUpub Connector
Deposited On:01 Sep 2022 07:34
Metadata Last Modified:01 Sep 2022 07:41

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