Genetic variability of Bipolaris sorokiniana isolates using URP-PCR

Abstract

Spot blotch disease, caused by Bipolaris sorokiniana, is one of the major diseases of wheat and is responsible for large losses of wheat crops worldwide. We used polymerase chain reaction (PCR) with universal rice primers (URP) for molecular characterization of 60 monosporic B. sorokiniana isolates from Brazil and other countries, and evaluated the diversity of the samples. PCR amplification generated 232 different DNA fragments ranging in size from 100 to 2018 bp. The primers URP-4R, URP-2R, and URP-1F generated greater numbers of amplified fragments (36, 30, and 25, respectively) from the single-spore isolates, and some diversity was observed among the isolates generated using these primers. The primers URP-2F, URP-6R, URP-17R, URP-30F, and URP-38F produced a pattern of monomorphic fragments and 73% of the isolates showed an average of 44 different DNA-amplified fragments. Primer URP-2F generated a 578-bp fragment that was common to 83.7% of the isolates; primer URP-6R generated a 548-bp fragment and primer URP-38F generated a 650-bp product that was common to 89.1% and 80% of the isolates, respectively. The URP-PCR primers provided important information about the genetic profiles of the monosporic cultures, which showed intraspecific variability among the monoconidial isolates and among the monosporic cultures that originated from the same polysporic strain. Our results indicate that URP's are sensitive and give reproducible results for assaying the genetic variability of B. sorokiniana.

Diversity of theFusariumpathogens associated with crown rot in the Huanghuai wheat‐growing region of China

“…Mann et al . () used URP‐PCR primers to evaluate the diversity of 60 monosporic Bipolaris sorokiniana isolates.…”

“…For instance, Cho et al (2012) found that the pathovars of Pseudomonas syringae could be divided into four clusters based on 63% similarity by URP-PCR using 2F, 9F and 17R primers. Mann et al (2014) used URP-PCR primers to evaluate the diversity of 60 monosporic Bipolaris sorokiniana isolates.…”

Molecular, morphological and pathogenic diversity ofSclerotinia sclerotiorumisolates from common bean (Phaseolus vulgaris) fields in Argentina

“…The URP markers proved to be a useful technique for the population analysis of S. sclerotiorum , revealing high polymorphism among the isolates analysed. The results suggest that, despite their dominant nature, URP markers complemented with mycelial compatibility assays are a useful tool in providing preliminary information on the genetic structure and reproductive behaviour of S. sclerotiorum , as reported for other pathogens (González et al ., ; Mann et al ., ).…”

“…Universal rice primers (URPs; Kang et al ., ) are dominant molecular markers that have been extensively used in fingerprinting studies for many pathogens (González et al ., ; Mann et al ., ). This technique has the advantage of being cost effective with no prior knowledge of the species genome needed for their use.…”

“…Moreover, URP markers are more reproducible than those of RAPD (random amplified polymorphic DNA) because stringent PCR conditions with high annealing temperatures and longer primers are employed (Kang et al ., ). Also, because they have multiple polymorphic loci, URP markers are suitable for the analysis of closely related individuals (Kang et al ., ; González et al ., ; Mann et al ., ). A previous study has considered the utility of URP markers for the analysis of S. sclerotiorum isolates; however, few markers were used (Manjunatha et al ., ).…”