Abstract
Gray mold disease inflicted by Botrytis cinerea is a serious menace responsible for significant economic loss worldwide. Due to its polyphagous nature, the pathogen has enthused inquisitiveness in researchers to unravel its complexity. Agrobacterium tumefaciens-mediated transformation was used to generate insertional mutants of Botrytis cinerea. A mutant (BCM-55) with disruption in a gene (BcDGAT2) that encodes for diacylglycerol O-acyl transferase 2 (DGAT2), showed enervated virulence on various hosts’ tissues. Enzyme DGAT2 is crucial in the final step of synthesis of triacylglycerol (TAG) that plays an important role in homeostasis of membrane and cellular processes. However, the role of DGAT2 has never been reported in a phytopathogenic fungus. In this study, BCM-55 was characterized to ascertain the role of DGAT2 in virulence of B. cinerea. The insertional mutant was defective in spore production and lacked sclerotia formation as a consequence of lower accumulation of TAG. A significant delay in spore germination in BCM-55 was accompanied with a low penetration potential. Hyphae of the mutant formed swollen endings with considerable impairment in penetration. Deletion of BcDGAT2 also led to increased sensitivity towards cell wall and membrane-disturbing agents. Furthermore, BCM-55 was deficient in the production of oxalic acid and showed lower activity of a cell wall-degrading enzyme, polygalacturonase. The role of BcDGAT2 in virulence was further confirmed by targeted deletion and complementation of the gene. The results insinuate a crucial role of BcDGAT2 in penetration and consequently virulence of B. cinerea. The study provides novel insights into plant–pathogen interactions that can be exploited to develop suitable disease management strategies.
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Acknowledgements
We thank Dr. Praveen Verma (NIPGR, New Delhi) for providing binary vector pBIF-EGFP and Prof. S. C. Bhatla (Department of Botany, University of Delhi) for his guidance in conducting experiments on lipid analysis. The authors gratefully acknowledge Rasmus John Normand Frandsen for his directions in carrying out targeted deletion of gene. Rupam Kapoor thankfully acknowledges the grant-in-aid provided by Science and Engineering Research Board (SERB), Government of India. Esha Sharma and Garima Anand are thankful to Department of Science and Technology and University Grants Commission for providing fellowship.
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Sharma, E., Tayal, P., Anand, G. et al. Functional analysis of diacylglycerol O-acyl transferase 2 gene to decipher its role in virulence of Botrytis cinerea . Curr Genet 64, 443–457 (2018). https://doi.org/10.1007/s00294-017-0752-1
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DOI: https://doi.org/10.1007/s00294-017-0752-1