PreprintArticleVersion 1Preserved in Portico This version is not peer-reviewed
Whole Genome Alignment Based Development of Molecular Marker for Detecting Leptosphaeria maculans and Leptosphaeria biglobosa, the Causal Agent of Blackleg Disease in Brassicas
Version 1
: Received: 12 October 2019 / Approved: 13 October 2019 / Online: 13 October 2019 (16:08:55 CEST)
How to cite:
Ferdous, M.J.; Hossain, M.R.; Jesse, D.M.I.; Park, J.; Jung, H.; Kim, H.; Nou, I. Whole Genome Alignment Based Development of Molecular Marker for Detecting Leptosphaeria maculans and Leptosphaeria biglobosa, the Causal Agent of Blackleg Disease in Brassicas. Preprints2019, 2019100147. https://doi.org/10.20944/preprints201910.0147.v1
Ferdous, M.J.; Hossain, M.R.; Jesse, D.M.I.; Park, J.; Jung, H.; Kim, H.; Nou, I. Whole Genome Alignment Based Development of Molecular Marker for Detecting Leptosphaeria maculans and Leptosphaeria biglobosa, the Causal Agent of Blackleg Disease in Brassicas. Preprints 2019, 2019100147. https://doi.org/10.20944/preprints201910.0147.v1
Ferdous, M.J.; Hossain, M.R.; Jesse, D.M.I.; Park, J.; Jung, H.; Kim, H.; Nou, I. Whole Genome Alignment Based Development of Molecular Marker for Detecting Leptosphaeria maculans and Leptosphaeria biglobosa, the Causal Agent of Blackleg Disease in Brassicas. Preprints2019, 2019100147. https://doi.org/10.20944/preprints201910.0147.v1
APA Style
Ferdous, M.J., Hossain, M.R., Jesse, D.M.I., Park, J., Jung, H., Kim, H., & Nou, I. (2019). Whole Genome Alignment Based Development of Molecular Marker for Detecting <em>Leptosphaeria maculans</em> and <em>Leptosphaeria biglobosa</em>, the Causal Agent of Blackleg Disease in Brassicas. Preprints. https://doi.org/10.20944/preprints201910.0147.v1
Chicago/Turabian Style
Ferdous, M.J., Hoy-Taek Kim and Ill-Sup Nou. 2019 "Whole Genome Alignment Based Development of Molecular Marker for Detecting <em>Leptosphaeria maculans</em> and <em>Leptosphaeria biglobosa</em>, the Causal Agent of Blackleg Disease in Brassicas" Preprints. https://doi.org/10.20944/preprints201910.0147.v1
Abstract
Background: Accurate diagnosis of the differentially aggressive fungus Leptosphaeria maculans and Leptosphaeria biglobosa causing Blackleg in crucifers is crucial. Available markers were designed decades ago which may become ineffective due to the ever evolving nature of the fungus, requiring the development of more precise molecular markers. Methods: The whole genomes of available isolates belonging to this two species were aligned using progressive MAUVE tool, species specific genomic regions were extracted and species specific primers were designed from the sequences that encode for effector proteins. Results: Three (Lm1, Lm2 and Lm5) and two (Lb3 and Lb3’) primer sets specifically detected the isolates of target species in PCR based assay, of which the primers Lm5 and Lb3’ were multiplexed for detection of Leptosphaeria maculans and Leptosphaeria biglobosa, generating PCR amplicons of 230 and 834 bp, respectively from a single PCR reaction. The markers were highly sensitive and were able to amplify target species from crude ‘pseudothecia and ascospores suspension’ without requiring DNA extraction. Conclusions: These markers, solitarily or in combination, designed from species specific genomic segments will serve as precise, sensitive and rapid detection of Leptosphaeria maculans and Leptosphaeria biglobosa species and will be helpful for surveillance, management and transboundary quarantine of the devastating disease.
Biology and Life Sciences, Biology and Biotechnology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
