title: Bcl-2△21 and Ac-DEVD-CHO Inhibit Death of Wheat Microspores.
abstract: Microspore cell death and low green plant production efficiency are an integral obstacle in the development of doubled haploid production in wheat. The aim of the current study was to determine the effect of anti-apoptotic recombinant human B-cell lymphoma-2 (Bcl-2△21) and caspase-3-inhibitor (Ac-DEVD-CHO) in microspore cell death in bread wheat cultivars AC Fielder and AC Andrew. Induction medium containing Bcl-2△21 and Ac-DEVD-CHO yielded a significantly higher number of viable microspores, embryo-like structures and total green plants in wheat cultivars AC Fielder and AC Andrew. Total peroxidase activity was lower in Bcl-2△21 treated microspore cultures at 96 h of treatment compared to control and Ac-DEVD-CHO. Electron paramagnetic resonance study of total microspore protein showed a different scavenging activity for Bcl-2△21 and Ac-DEVD-CHO. Bcl-2△21 scavenged approximately 50% hydroxyl radical (HO•) formed, whereas Ac-DEVD-CHO scavenged approximately 20% of HO•. Conversely, reduced caspase-3-like activities were detected in the presence of Bcl-2△21 and Ac-DEVD-CHO, supporting the involvement of Bcl-2△21 and Ac-DEVD-CHO in increasing microspore viability by reducing oxidative stress and caspase-3-like activity. Our results indicate that Bcl-2△21 and Ac-DEVD-CHO protects cells from cell death following different pathways. Bcl-2△21 prevents cell damage by detoxifying HO•and suppressing caspase-3-like activity, while Ac-DEVD-CHO inhibits the cell death pathways by modulating caspase-like activity.
authors.affiliation: Cereal Biotechnology, Agriculture and Agri-Food Canada, LethbridgeAB, Canada; Department of Plant Anatomy and Cytology, Faculty of Biology and Environmental Protection, University of SilesiaKatowice, Poland.
authors.forename: Rakesh K
authors.initials: RK
authors.lastname: Sinha
authors.affiliation: Department of Biophysics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University Olomouc, Czechia.
authors.forename: Pavel
authors.initials: P
authors.lastname: Pospíšil
authors.affiliation: Cereal Biotechnology, Agriculture and Agri-Food Canada, Lethbridge AB, Canada.
authors.forename: Priti
authors.initials: P
authors.lastname: Maheshwari
authors.affiliation: Cereal Biotechnology, Agriculture and Agri-Food Canada, Lethbridge AB, Canada.
authors.forename: François
authors.initials: F
authors.lastname: Eudes
keywords: Bcl-2△21; caspase-3; cell death; embryogenesis; hydroxyl radicals; microspore