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United States Department of Agriculture

Agricultural Research Service

Research Project: Detection and Identification of Soil-borne Plant Pathogens and Mitigation of Soil-borne Diseases

Location: Floral and Nursery Plants Research Unit

Title: Development of SCAR markers and UP-PCR cross-hybridization method for specific detection of four major subgroups of Rhizoctonia from infected turfgrasses

Authors
item Amaradasa, Bimal -
item Horvath, Brandon -
item Lakshman, Dilip

Submitted to: Mycologia
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 6, 2013
Publication Date: May 23, 2013
Citation: Amaradasa, B.S., Horvath, B.J., Lakshman, D.K. 2013. Development of SCAR markers and UP-PCR cross-hybridization method for specific detection of brow patch pathogens from infected turfgrass. Lecture. 106(1):163-172.

Technical Abstract: Several species and hyphal anastomosis groups (AG) of Rhizoctonia solani (sensu lato) cause brown patch diseases of turfgrasses. Conventional methods of identification of Rhizoctonia pathogens are time consuming and often inaccurate. A rapid identification assay for Waitea circinata (anamorph: Rhizoctonia spp.) varieties zeae and circinata causing patch diseases was developed based on the universally primed PCR (UP-PCR) products cross-blot hybridization with non-radioactive probes. Isolates within a W. circinata variety cross-hybridized strongly while non-homologous isolates did not cross-hybridize or did so weakly. Closely related W. circinata varieties zeae and circinata were clearly distinguished using this assay. In addition, sequence-characterized amplified region (SCAR) markers were developed from UP-PCR products to identify turf pathogenic isolates of Thanatephorus cucumeris (anamorph: R. solani) AG 1-IB and AG 2-2IIIB. The developed SCAR markers could distinguish isolates of AG 1-IB or AG 2-2IIIB groups and did not amplify any product from genomic DNA of non-target isolates of Rhizoctonia. The specific primers were sensitive and unique enough to produce a PCR band from total DNA of diseased turfgrasses infected with either AG 1-IB or AG 2-2IIIB.

Last Modified: 10/25/2014
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