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In vitro regeneration of two Populus hybrid clones. The role of pectin domains in cell processes underlying shoot organogenesis induction

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Biologia Plantarum

Abstract

An efficient plant regeneration protocol has been established for two commercial Populus hybrid clones, MC (Populus × euramericana) and UNAL (Populus × interamericana). The culture of internode segments on Murashige and Skoog (MS) medium with 0.5 μM α-naphthalene acetic acid (NAA) and 4 μM N6-benzyladenine for 7 weeks (2 weeks in absence of activated charcoal and 5 weeks in its presence) resulted in the highest frequency of shoot regeneration (100 % for MC and 82 % for UNAL). All regenerated shoots longer than 2 cm rooted on half-strength MS medium, independent of the addition of 0.1 μM NAA. Nevertheless, shoots developed better-formed roots in NAA-free medium, which had a positive effect on the acclimatization of plants. In order to know the cellular processes underlying in vitro shoot organogenesis, a histological study was made in UNAL internode-explants. Results revealed that in vitro culture caused swelling around the cut-off zones in all explants, but only those undergoing organogenesis formed proliferation centers under subepidermal cells, which led to formation of bud primordia. Moreover, in vivo tissues and explants with different in vitro response showed different immunolabelling patterns when they were treated with fluorescentmonoclonal antibodies directed to several pectin-polysaccharides of the cell wall. Results allow us to assign a predominant role of homogalacturonan with a low degree of methyl-esterification in the initiation of bud primordia, a role of β-1,4-D-galactan side chains of rhamnogalacturonan-I in the cellular differentiation, ra ole of α-1,5-L-arabinan side chains of rhamnogalacturonan-I and of homogalacturonan with a high degree of methyl-esterification in cell division and growth.

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Abbreviations

AC:

activated charcoal

BA:

N6-benzyladenine

HG:

homogalacturonan

NAA:

a-naphthalene acetic acid

PBS:

phosphate-buffered saline

PFD:

photon flux density

PGR:

plant growth regulator

RG-I:

rhamnogalacturonan-I.

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Correspondence to M. L. Centeno.

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Acknowledgment: The authors are thankful to the Head Department of Plant Engineering and Sciences of the University of León for providing all the resources to carry out the research work. The assistance in providing poplar clones MC and UNAL by the National Centre of Forest Genetic Resources is also highly acknowledged. We thank Rafael Calvo Sanz for his help with this manuscript and Darren Arnott for the linguistic revision. Finally, we would like to thank Santiago Michavila for his collaboration.

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García-Angulo, P., Villar, I., Giner-Robles, L. et al. In vitro regeneration of two Populus hybrid clones. The role of pectin domains in cell processes underlying shoot organogenesis induction. Biol Plant 62, 763–774 (2018). https://doi.org/10.1007/s10535-018-0819-y

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  • DOI: https://doi.org/10.1007/s10535-018-0819-y

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