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. 2017 Aug 24;12(8):e0181785.
doi: 10.1371/journal.pone.0181785. eCollection 2017.

Nitrogen uptake and assimilation in proliferating embryogenic cultures of Norway spruce-Investigating the specific role of glutamine

Johanna Carlsson  1   2 , Henrik Svennerstam  1   3 , Thomas Moritz  1 , Ulrika Egertsdotter  1   4 , Ulrika Ganeteg  1
Affiliations

Nitrogen uptake and assimilation in proliferating embryogenic cultures of Norway spruce-Investigating the specific role of glutamine

Johanna Carlsson et al. PLoS One. .

Erratum in

Abstract

Somatic embryogenesis is an in vitro system employed for plant propagation and the study of embryo development. Nitrogen is essential for plant growth and development and, hence, the production of healthy embryos during somatic embryogenesis. Glutamine has been shown to increase plant biomass in many in vitro applications, including somatic embryogenesis. However, several aspects of nitrogen nutrition during somatic embryogenesis remain unclear. Therefore, we investigated the uptake and assimilation of nitrogen in Norway spruce pro-embryogenic masses to elucidate some of these aspects. In our study, addition of glutamine had a more positive effect on growth than inorganic nitrogen. The nitrogen uptake appeared to be regulated, with a strong preference for glutamine; 67% of the assimilated nitrogen in the free amino acid pool originated from glutamine-nitrogen. Glutamine addition also relieved the apparently limited metabolism (as evidenced by the low concentration of free amino acids) of pro-embryogenic masses grown on inorganic nitrogen only. The unusually high alanine concentration in the presence of glutamine, suggests that alanine biosynthesis was involved in alleviating these constraints. These findings inspire further studies of nitrogen nutrition during the somatic embryogenesis process; identifying the mechanism(s) that govern glutamine enhancement of pro-embryogenic masses growth is especially important in this regard.

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Conflict of interest statement

Competing Interests: One author is affiliated with Svenska Skogsplantor AB, Seed Production, Lagan, Sweden. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1. PEM biomass after two weeks of growth on three different experimental media (PM#1–3), for cell lines 11:08:59 and 11:12:02.
(A–B) Pictures of the PEMs. (C–D) Biomass. (E–F) Dried biomass. (G–H) Ratio of FW/DW. Each bar represents a mean ± SE; n = 10. Different letters above the bars in respective panels indicate significant differences between the treatments at P<0.05 (Tukey’s test).
Fig 2
Fig 2. Total free AA pool concentration for PEMs grown on three different experimental media (PM#1–3).
(A) Cell lines 11:08:59, (B) 11:12:02. Each bar represents a mean ± SE; n = 5. Different letters above the bars in respective panels indicate significant differences between the treatments at P<0.05 (Tukey’s test).
Fig 3
Fig 3. Profile of AA concentrations in PEMs grown on medium PM#3.
Cell line 11:08:59 (black bars) and 11:12:02 (white bars). Each bar represents a mean ± SE; n = 5.
Fig 4
Fig 4. Fraction and amount of N from each N source, NH4+, NO3- and L-Gln detected in the PEMs (mean mg N ± SE; n = 10).
Fig 5
Fig 5. Assimilation of N into the total free AA pool in PEMs grown on PM#3.
Fraction and amount of N from each N source, NH4+, NO3- and L-Gln (mean μg N ± SE; n = 9–10).
See this image and copyright information in PMC

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