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Pinus sylvestris L. mature seeds megagamethophytes cultured in vitro. Influence of the genotype and the growth regulators content in nutrient medium on the initiation of callus formation

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Abstract

Despite the significant progress in forest biotechnology Scots pine still remains difficult for mass propagation via somatic embryogenesis (SE) due to the influence of many factors, such as genotype and location of the donor plant, the composition of the nutrient medium, including the concentration of growth regulators, etc. Mature seeds were collected from Pinus sylvestris trees growing in the northern (Loukhsky district, Engozerskoye district forestry, natural phytocoenosis) and middle (Prionezhsky district, Petrozavodsk seed orchard) boreal subzone in Karelia (Russia). The influence of various growth regulators content in the composition of the DCR medium on the initiation of callus formation from mature seeds megagametophytes of both habitats was analyzed. Cytogenetic analysis of explants (root meristem of seedlings) showed that the frequency of mitosis pathologies is 4 times, and the proportion of cells with micronuclei is 6 times higher in P. sylvestris plants from a natural phytocoenosis than from Petrozavodsk seed orchard. Nevertheless explants collected from trees growing in the northern boreal subzone more often formed a cell culture (initiation frequency 13.4%) than megagametophytes from seed orchard (8.83%). The most active callus formation was registered on medium containing (1) 9 µM benzyladenine and 13.6 µM 2,4-dichlorophenoxyacetic acid; (2) 9 µM benzyladenine and 2.7 µM 1-naphthylacetic acid.

Key Message

It was shown that the initiation of callus formation depends both on the content of plant growth regulators in the medium and on the explant donor plant.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

SE:

Somatic Embryogenesis

2.4-D:

2,4-Dichlorophenoxyacetic acid

BA:

6-Benzyladenine

NAA:

1-Naphthylacetic acid

IAA:

Indolylacetic acid

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Funding

The study was carried out under state order to the Karelian Research Centre of the Russian Academy of Sciences and under the world-class scientific and educational centre called Russian Arctic: New Materials, Technologies, and Research Methods.

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Authors and Affiliations

  1. Department of Multidisciplinary Scientific Research of Karelian Research Centre of the Russian Academy of Sciences, 11 Pushkinskaya St, Karelia, Petrozavodsk, Russia, 185910

    Olga V. Chirva, Roman V. Ignatenko & Maria A. Ershova

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  1. Olga V. Chirva
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  2. Roman V. Ignatenko
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  3. Maria A. Ershova
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by OVC, RVI, and MAE. The first draft of the manuscript was written by OVC and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Olga V. Chirva.

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Communicated by Paloma Moncaleán.

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Chirva, O.V., Ignatenko, R.V. & Ershova, M.A. Pinus sylvestris L. mature seeds megagamethophytes cultured in vitro. Influence of the genotype and the growth regulators content in nutrient medium on the initiation of callus formation. Plant Cell Tiss Organ Cult 152, 299–308 (2023). https://doi.org/10.1007/s11240-022-02404-3

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