Abstract
Plant regeneration from reproductive organs is a poorly understood route in passion fruit. The present study provides a morpho-hystological description of somatic embryogenesis from anther culture in Passiflora gibertii N. E. Br., as well as a phytochemical profile and genetic analysis of ploidy in the generated embryos. Flower buds were collected during the pre-anthesis period. Murashige and Skoog culture medium was supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzyladenine. Embryogenic calluses were collected at different ages for histological, biochemical, and genetic evaluation of their ploidy. Anatomical characterization revealed the cell layers of the epidermal tissue involved in morphogenic events. Specifically, regions of intense cell division led to embryo formation and its subsequent regeneration. Moreover, as shown by hormonal and secondary compounds’ profiles, these regions were characterized by the presence of reserve compounds and higher levels of proteins and pectins. Finally, flow cytometry confirmed the diploid nature of the embryos. The present results define a pioneering study of anthers in P. gibertii N. E. Brown, and could serve as a blueprint for future in vitro regeneration protocols.
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The authors are grateful to Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the fellowship offered to the lead author during his PhD studies. We would like to thank NuBiomol/UFV for hormonal analysis and Editage (https://www.editage.com) for English language editing.
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This work was supported by the Brazilian agencies Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG, Belo Horizonte, MG, Brazil; Grants APQ-01131-15 and APQ-00772-19), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasília, DF, Brazil; Grant Finance Code 001), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brasília, DF, Brazil; Grants 435488/2016-3 and 420913/2018-1).
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WSS, ADK, and WCO designed the experiments. WSS and ADK established the cultures. ACFC and ADK performed anatomical and light microscopy analyses, as well as hormonal and flavonoid. EMM and LFV performed flow cytometry analysis. WSS, DIR, CHB, LFV, CHB, and WCO wrote the paper.
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dos Santos Soares, W., Bruckner, C.H., da Cruz, A.C.F. et al. Shining light on anther culture, a poorly understood regeneration route in passion fruit (Passiflora gibertii N. E. Brown): histological, hormonal, and phytochemical aspects. In Vitro Cell.Dev.Biol.-Plant 57, 998–1008 (2021). https://doi.org/10.1007/s11627-021-10179-z
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DOI: https://doi.org/10.1007/s11627-021-10179-z