Insights into the early stage of Pinus nigra Arn. somatic embryogenesis using discovery proteomics

Katarína Klubicová; Lubica Uvácková; Maksym Danchenko; Peter Nemecek; Ludovít Skultéty; Ján Salaj; Terézia Salaj

doi:10.1016/j.jprot.2017.05.013

Insights into the early stage of Pinus nigra Arn. somatic embryogenesis using discovery proteomics

J Proteomics. 2017 Oct 3:169:99-111. doi: 10.1016/j.jprot.2017.05.013. Epub 2017 May 17.

Authors

Katarína Klubicová¹, Lubica Uvácková², Maksym Danchenko³, Peter Nemecek⁴, Ludovít Skultéty³, Ján Salaj⁵, Terézia Salaj⁵

Affiliations

¹ Institute of Plant Genetics and Biotechnology, Plant Science and Biodiversity Center, Slovak Academy of Sciences, Akademická 2, P.O. Box 39A, 950 07 Nitra, Slovakia. Electronic address: katarina.klubicova@savba.sk.
² Institute of Plant Genetics and Biotechnology, Plant Science and Biodiversity Center, Slovak Academy of Sciences, Akademická 2, P.O. Box 39A, 950 07 Nitra, Slovakia; Department of Biology, Faculty of Natural Sciences, University of SS. Cyril and Methodius, Nám. J. Herdu 2, 917 01 Trnava, Slovakia.
³ Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Dúbravská cesta 9, 845 05 Bratislava, Slovakia.
⁴ Department of Chemistry, Faculty of Natural Sciences, University of SS. Cyril and Methodius, Nám. J. Herdu 2, 917 01 Trnava, Slovakia.
⁵ Institute of Plant Genetics and Biotechnology, Plant Science and Biodiversity Center, Slovak Academy of Sciences, Akademická 2, P.O. Box 39A, 950 07 Nitra, Slovakia.

PMID: 28526530
DOI: 10.1016/j.jprot.2017.05.013

Abstract

The somatic embryogenesis in conifers represents a suitable model of plant regeneration system facilitating studies of fundamental aspects of an early development as well as in vitro micropropagation. The aim of our study was to deeper understand the somatic embryogenesis in the conifer tree Pinus nigra Arn. Comparative proteomic analysis based on 2D-PAGE in 1) proliferating embryogenic tissues (E) initiated from immature zygotic embryos, 2) non-embryogenic calli (NEC) initiated from cotyledons of somatic seedlings of the same genotypes, 3) embryogenic tissues that lost the maturation capacity (E-L) of two cell lines (E362, E366). Investigated pine tissues showed distinct structural features. The 24 protein spots were altered in both cell lines in comparison of embryogenic and non-embryogenic tissues. These proteins are involved in disease and defence mechanism, energy metabolism and biosynthesis of cell wall components. Two of three protein spots detected only in embryogenic form of both cell lines are similar to water deficit inducible protein LP3, the third remains uncharacterised. The loss of the maturation capacity was accompanied by changes in 35 and 38 protein spots in 362 and 366 cell lines, respectively. Only two of them were altered in both cell lines, suggesting non-uniform process of ageing.

Biological significance: Somatic embryogenesis in conifers represents an experimental system for the study of early plant development as well as a biotechnological tool for large-scale micropropagation. The obtained results give a new insight into the process of somatic embryogenesis of a conifer Pinus nigra Arn. by revealing differences at proteomic levels among in vitro cultured tissues characterised by different embryogenic potential. Microscopic investigations have also shown differences in the structural organisation of studied tissues.

Keywords: 2-D gel electrophoresis; Cell wall biosynthesis; Conifers; Embryogenic tissue; Loss of maturation capacity; Non-embryogenic callus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Electrophoresis, Gel, Two-Dimensional
Pinus / embryology*
Pinus / growth & development
Plant Proteins / metabolism
Plant Somatic Embryogenesis Techniques / methods
Proteomics / methods*

Substances

Plant Proteins