Tilletia walkeri, a New Species on Lolium multiflorum and L. perenne

Castlebury, Lisa A.; Carris, L. M.

doi:10.2307/3761200

Cited by 40 publications

(24 citation statements)

References 20 publications

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“…T. walkeri shares >99 % identity with T. indica, but is more commonly found in ryegrass and does not cause Karnal bunt (Levy et al 2001). Indeed, PCR amplification has resulted in mis-identification of the disease using published primers, mistaking T. walkeri as T. indica (Castlebury and Carris 1999;Ferreira et al 1996;Pimentel et al 1998;Smith et al 1996). Several strategies have been designed to distinguish T. indica from other Tilletia species when using PCR-based technique.…”

Section: Introductionmentioning

confidence: 96%

Rapid and specific detection of Tilletia indica using loop-mediated isothermal DNA amplification

Gao

Tan

Zhu

2016

Australasian Plant Pathol.

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Karnal bunt in wheat is caused by the fungus Tilletia indica. It is a quarantine disease, and therefore its timely and specific detection is important. Current detection protocols involve DNA amplification by polymerase chain reaction (PCR). However, this technique has encountered specificity issues due to the high DNA homology of T. indica with other Tilletia species, in particular T. walkeri. Here, we report the specific and rapid detection of T. indica DNA using the loop-mediated isothermal amplification (LAMP) at 62°C. Alignment of the mitochondrial DNA of T. indica and T. walkeri has revealed four major unique regions in T. indica. Six LAMP primers designed in one of these unique regions were able to amplify T. indica DNA. The amplification could be completed in 30 min and was nearly as sensitive as conventional PCR. The amplification was found to be highly specific to T. indica DNA during the screening of 17 isolates of T. indica, T. walkeri, T. horrida, T. ehrhartae and T. caries. The use of the fluorescent chemical calcein has enabled endpoint detection with the naked eye. This method, with its specificity, rapidity and simple visualization, is well suited for the detection of this important disease in wheat.

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Section: Introductionmentioning

confidence: 96%

Rapid and specific detection of Tilletia indica using loop-mediated isothermal DNA amplification

Gao

Tan

Zhu

2016

Australasian Plant Pathol.

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“…are grown in rotation with Tr. aestivum (Castlebury & Carris, 1999;Pascoe, Priest, Shivas, & Cunnington, 2005). Microscopic method of detection lacks consistency, accuracy and sensitivity, which results in frequent misidentifications of KB (Barry & Castlebury, 1999).…”

Section: Generation Of Diagnostic Anti-teliospore Antibodies For Devementioning

confidence: 98%

Generation of diagnostic anti-teliospore antibodies for development of specific immunodiagnostic format for detection of Karnal bunt (Tilletia indica) of wheat

Singh

Pandey

et al. 2014

Food and Agricultural Immunology

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Rabbit antisera were generated against four preparations of teliospores of Tilletia indica the fungus responsible for Karnal bunt in wheat having titre ranging from 1:2500 to 1:10,000. The suitability of four solubilising agents (I-IV) used for extraction of teliospores/spore proteins was assessed, highest protein extractability was observed with solubilising agent I (0.5% sodium dodecyl sulphate). The maximum immunoreactivity in enzyme linked immunosorbent assay (ELISA) was observed with the teliosporic protein(s) extracted using solubilising agent I followed by agent II, IV and III. Further, the results of western blotting with proteins from different fungal pathogens using anti-intact teliospores antibody showed one unique immunoreactive band of 28 kDa with teliosporic protein of T. indica only. The potential diagnostic antibody was generated against the characterised protein (28 kDa) of teliospore wall's antigen of T. indica. The developed diagnostic antibody and solubilising agent I could therefore be employed for development of specific and rapid immunodiagnostic format for detection of Karnal bunt.

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“…Such a method must meet three requirements: (a) a detection method sensitive enough to detect one teliospore in 50g sample; (b) an extraction method that is quick and cost effective in the evaluation of a large number of samples; and (c) compatibility with available biochemical and molecular methods for identification of T. indica (Bonde, Pesterson, Dowler, & May, 1989;Castlebury & Carris, 1999;Ferreira, Tooley, Hatziloukas, Castro, & Schaad, 1996;Frederick et al 1998;Luster, Bonde, Peterson, Hack, & Russo, 1995;Matsumoto & Bell, 1989;McDonald, Wong, Kristjansson, & White, 1999;Smith, Peterson, Beck, Schaad, & Bonde, 1996).…”

Section: Requirements Of Molecular Diagnosticsmentioning

confidence: 99%

Application of molecular and immuno-diagnostic tools for detection, surveillance and quarantine regulation of Karnal bunt (Tilletia indica) of wheat

Kumar

Singh

Kumar

et al. 2008

Food and Agricultural Immunology

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Disease diagnostics and pathogen detection, a primary component of any crop management programme, is also helpful in monitoring sanitary and phyto-sanitary measures to seed quality. Molecular diagnostics based on immunological and DNA techniques can provide an efficient way for disease surveillance and disease forecasting. Karnal bunt (KB) of wheat has become a major disease of economic importance and an important issue in the international wheat trade. Due to its seed borne, soil borne and air borne nature, KB can be diagnosed in three pathogenic forms of pathogens viz. teliospores, sporidial and mycelial forms which occur in infectious or proliferative entities at different stages of the disease cycle. Suitable sampling and extraction procedures can improve the detection limits of modern molecular diagnostic methods used to test KB. For the extraction of teliospores from both infected and infested seeds, highly efficacious detergent washing techniques have been developed and extracted pathogenic entities were diagnosed by involving several species of specific primers and immuno-chemical reagents. Nucleic acid and antibody based techniques have been developed for the detection of KB pathogen in different forms. These techniques can also be successfully employed for differential diagnosis, disease surveillance of seed borne pathogens of quarantine importance and determination of teliospore load in wheat seeds. These tests can also be designed to assist in the trade of wheat, development of disease forecasting models and molecular and immuno-pathotyping of Tilletia indica isolates collected from different agro-climatic regions. Moreover, these formats could also be employed for molecular characterization of disease determinants and in comprehending their role in molecular basis of host-pathogen interaction. The present paper highlights the use of the principle of molecular diagnostics in exerting disease surveillance and quality assurances of seeds.

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Tilletia walkeri, a New Species on Lolium multiflorum and L. perenne

Cited by 40 publications

References 20 publications

Rapid and specific detection of Tilletia indica using loop-mediated isothermal DNA amplification

Rapid and specific detection of Tilletia indica using loop-mediated isothermal DNA amplification

Generation of diagnostic anti-teliospore antibodies for development of specific immunodiagnostic format for detection of Karnal bunt (Tilletia indica) of wheat

Application of molecular and immuno-diagnostic tools for detection, surveillance and quarantine regulation of Karnal bunt (Tilletia indica) of wheat

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