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Cellular and morphological changes during leaf explant dedifferentiation and plant regeneration of Tagetes erecta

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Abstract

Plant tissue culture allows the maintenance of species of interest under controlled conditions. There have been few studies related to the morphological changes that take place during callus formation and redifferentiation processes. The aim of this work was to analyze the morphological changes during dedifferentiation and redifferentiation processes in leaf explants of marigold (Tagetes erecta L.) using transmission electron microscopy, digital image analysis and concepts of fractal dimension. Leaf explants were sown in MS medium supplemented with 2.0 mg L−1 of benzyladenine and 2.0 mg L−1 of 2,4-dichlorophenoxyacetic acid to induce dedifferentiation (callus formation). The redifferentiation process (bud formation) was obtained with explants sown in MS medium supplemented with 3.0 mg L−1 indoleacetic acid and 0.5 mg L−1 benzyladenine. During the first 8 days, the dedifferentiation process showed gradual changes in terms of the area and perimeter of the explants. The fractal dimension of the perimeter corresponding to the projected image of the calli changed from smooth to rough at day 20. These features indicate variations in tissue morphometry as a result of the callus formation process and further formation of cellular aggregates. Structural observations during the dedifferentiation process showed changes in shape and cellular organization as callus formation became evident. Round-shaped cells were observed in the callus, with few organelles and a low degree of differentiation. Indirect organogenesis was achieved over a period of 10 days; in addition, plantlets developed from the obtained buds in 20 days in growth regulator-free MS medium.

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Acknowledgements

The authors are thankful to the Secretaría de Investigación y Posgrado of the Instituto Politécnico Nacional (Grant No. 20180804) of México for the support granted for the accomplishment of this work.

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Authors and Affiliations

  1. Instituto Politécnico Nacional, CEPROBI, Carretera Yautepec-Jojutla km. 6, Calle CEPROBI 8, Col. San Isidro, C.P. 62731, Yautepec, Morelos, México

    Pablo Emilio Vanegas-Espinoza, Crescencio Bazaldúa & Alma Angélica Del Villar-Martínez

  2. Instituto Tecnológico José Mario Molina Pasquel y Henriquez, Campus Tamazula de Gordiano, Carretera Tamazula Santa Rosa No. 329, La Huerta, México

    Leticia Betsaida Ríos-Salomé

  3. Centro de Investigación y de Estudios Avanzados-IPN, Campus Irapuato, Guanajuato, México

    Octavio Paredes-López

  4. Sensient Colors - Latin America, Rodolfo Patrón No. 12 Parque, Ind. Lerma., CP 52000, Lerma, Edo. de México, México

    Gustavo Adolfo Camelo-Méndez

Authors
  1. Pablo Emilio Vanegas-Espinoza
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  2. Crescencio Bazaldúa
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  3. Leticia Betsaida Ríos-Salomé
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  4. Octavio Paredes-López
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  5. Gustavo Adolfo Camelo-Méndez
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  6. Alma Angélica Del Villar-Martínez
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Contributions

This work was designed and supervised by AAVM and OPL. Sample collection and laboratory work were conducted by LBRS and CB, and analysis of results was performed by PEVE and GACM.

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Correspondence to Alma Angélica Del Villar-Martínez.

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Communicated by Tae-Ho Han, Ph.D.

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Vanegas-Espinoza, P.E., Bazaldúa, C., Ríos-Salomé, L.B. et al. Cellular and morphological changes during leaf explant dedifferentiation and plant regeneration of Tagetes erecta. Hortic. Environ. Biotechnol. 61, 407–414 (2020). https://doi.org/10.1007/s13580-019-00210-z

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  • DOI: https://doi.org/10.1007/s13580-019-00210-z

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