Trials of direct detection and identification of Rhizoctonia solani AG 1 and AG 2 subgroups using specifically primed PCR analysis

Abstract

Specifically primed polymerase chain reaction (PCR) analysis was used for direct detection and identification of Rhizoctonia solani isolates belonging to AG 1 subgroups (IA, IB, and IC) and AG 2 subgroups (2-1 and 2-2). A rapid DNA extraction method with a solution of sodium hydroxide was conducted to extract PCR templates. PCRspecific primer sets for each group were designed from sequences in the regions of the 28S ribosomal DNA of this fungus. The results of specifically primed PCR analysis showed that AG 1-IA, AG 1-IB, AG 1-IC, AG 2-1, and AG 2-2 primers sets contributed detection from the same AG isolates and could escape detection from different AG isolates at a high level of frequency. In this experiment, we suggested that our synthesized primer sets might provide a method for the direct detection and identification of AGs of R. solani.

Genetic Structure and Aggressiveness of Rhizoctonia solani AG1‐IA, the Cause of Sheath Blight of Rice in Southern China

“…The identity of the anastomosis group of R. solani AG1‐IA isolates was determined by PCR amplification of part of the 28S ribosomal DNA (rDNA) region using specific primers: AG‐common primer (forward) (5′‐CTCAAACAGGCATGCTC‐3′) and AG1‐IA specific primer (reverse) (5′‐CAGCAATAGTTGGTGGA‐3′) (Matsumoto ;. DNA extraction was achieved according to the cetyltrimethylammonium bromide (CTAB) method, as described by Carling et al.…”

The advancing identification and classification of Rhizoctonia spp. using molecular and biotechnological methods compared with the classical anastomosis grouping

“…Real-time PCR assays using TaqMan probes have been described for AG 3-PT from soil and on potato tubers (Lee et al 2002). Specific PCR primers designed from the 28S rDNA region were successfully used for the identification of MNR subgroups of AG 1 (-IA, -IB, and -IC) and AG 2 (2-1 and 2-2) (Matsumoto 2002).…”

Pathogenicity of Rhizoctonia solani AG 1-IB on common weeds in Meghalaya

“…Experiments related to "Pathogenicity of R. solani AG 1-IB on common weeds in Meghalaya" were conducted at ICAR Research Complex for NEH Region, Umiam, Meghalaya during 2016-2017. Infected soybean samples exhibiting web blight symptoms were collected from experimental plots of ICAR, Umiam and isolation was done following the method as described in Saveinai (2016) PCR was used for amplification of extracted DNA with specific primers for AG 1-IA and IB (Matsumoto 2002 andSayler andYang 2007). Both positive and negative controls were also used.The DNA extraction, PCR (with minor modifications), gel documentation and identification was done following the procedure described by Mahendra et al (2016).One isolate (PPJ3) was also amplified and Highly susceptible weeds identified in this study should be avoided for mulching purpose since this will increase the inoculum load of this pathogen sequenced by using universal primers for ITS region (ITS 5 and 4) (White et al 1990) and the sequence was deposited in Genbank.…”