Sheikh, I. | bioRxiv Science

bioRxiv2025

Development and validation of KASP markers for a novel powdery mildew resistance gene in wheat using BSR-Seq analysis

Wheat powdery mildew occurs as a devastating infection caused by the pathogenic fungi Blumeeria graminis f. sp. tritici (Bgt). The management of disease becomes much more effective when host resistance methods are employed. The WL711-2U/2B line served as the introgressed line to identify genes linked with powdery mildew resistance. This allowed us to look into the potential resistance components in response to powdery mildew infection. The resistance status of the line towards powdery mildew developed from a single dominant gene named PmAT. Bulked segregant RNA-Seq (BSR-Seq) together with previous QTL-Seq SNP markers helped identify strong candidate regions in chromosome arm 2B extending from 531114848 bp to 530235618 bp. The LOD score from this region reached 72.4 with a PVE% of 64.7, thus proving its main role in resistance. The physical position was subsequently locked up, which was different from the previously identified genes on the same chromosome arm in its position, suggesting that it is most likely a new Pm gene. The QTL interval contained five potential genes that we identified during this research. The five candidate genes directs the protein production, which performs functions linked to the pentatricopeptide repeat family as well as B-box-type zinc finger domain, P-loop-containing nucleoside triphosphate hydrolase, and plant peroxidase. HighlightThe identification of a new wheat powdery mildew resistance gene (PmAT) using a combination of BSR-Seq and QTL-Seq analysis, which located five candidate genes specifically on chromosome 2B.

genetics↗

bioRxiv2020

Whole genome sequencing for diagnosis of neurological repeat expansion disorders

BackgroundRepeat expansion (RE) disorders affect ~1 in 3000 individuals and are clinically heterogeneous diseases caused by expansions of short tandem DNA repeats. Genetic testing is often locus-specific, resulting in under diagnosis of atypical clinical presentations, especially in paediatric patients without a prior positive family history. Whole genome sequencing (WGS) is emerging as a first-line test for rare genetic disorders, but until recently REs were thought to be undetectable by this approach. MethodsWGS pipelines for RE disorder detection were deployed by the 100,000 Genomes Project and Illumina Clinical Services Laboratory. Performance was retrospectively assessed across the 13 most common neurological RE loci using 793 samples with prior orthogonal testing (182 with expanded alleles and 611 with alleles within normal size) and prospectively interrogated in 13,331 patients with suspected genetic neurological disorders. FindingsWGS RE detection showed minimum 97{middle dot}3% sensitivity and 99{middle dot}6% specificity across all 13 disease-associated loci. Applying the pipeline to patients from the 100,000 Genomes Project identified pathogenic repeat expansions which were confirmed in 69 patients, including seven paediatric patients with no reported family history of RE disorders, with a 0.09% false positive rate. InterpretationWe show here for the first time that WGS enables the detection of causative repeat expansions with high sensitivity and specificity, and that it can be used to resolve previously undiagnosed neurological disorders. This includes children with no prior suspicion of a RE disorder. These findings are leading to diagnostic implementation of this analytical pipeline in the NHS Genomic Medicine Centres in England. FundingMedical Research Council, Department of Health and Social Care, National Health Service England, National Institute for Health Research, Illumina Inc

genomics↗