AUTHOR=Kashyap Prem Lal , Kumar Sudheer , Kumar Ravi Shekhar , Sharma Anju , Khanna Annie , Raj Shubham , Jasrotia Poonam , Singh Gyanendra 

TITLE=Molecular diagnostic assay for pre-harvest detection of Tilletia indica infection in wheat plants

JOURNAL=Frontiers in Microbiology

VOLUME=14

YEAR=2023

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2023.1291000

DOI=10.3389/fmicb.2023.1291000

ISSN=1664-302X

ABSTRACT=<p>The current study describes a new diagnostic method for the rapid and accurate detection of <italic>Tilletia indica</italic>, the pathogen accountable for causing Karnal bunt (KB) disease in wheat. This method uses quantitative real-time polymerase chain reaction (qPCR) and a primer set derived from glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene of <italic>T. indica</italic> to identify the presence of the pathogen. The qPCR assay using this primer set was found highly sensitive, with a limit of detection (LOD) value of 4 pg of <italic>T. indica</italic> DNA. This level of sensitivity allows for the detection of the pathogen even in cases of different growth stages of wheat, where no visible symptoms of infection on the wheat plants can be seen by naked eyes. The study also validated the qPCR assay on ten different wheat cultivars. Overall, this study presents a valuable molecular tool for rapid, specific and sensitive detection of KB fungus in wheat host. This method has practical applications in disease management, screening of wheat genotypes against KB and can aid in the development of strategies to mitigate the impact of Karnal bunt disease on wheat production.</p>