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Comprehensive analysis of in vitro to ex vitro transition of tissue cultured potato plantlets grown with or without sucrose using metabolic profiling technique

Abstract

This study elucidated the effect of exogenous sucrose on growth parameters and metabolic changes during the in vitro rooting (InVR) and the ex vitro acclimatization (ExVA) stages of potato (Solanum tuberosum L.). During InVR stage, plantlets were cultured on MS medium with 3 % (S+) or without (S−) sucrose, and were then acclimatized under the same ExVA condition. In InVR stage, S+ increased photosynthetic capacity (Amax) and dry matter percentage. Yet, no significant differences in the other growth parameters have been observed. During acclimatization, Amax and respiration were higher in ExVA compared to InVR plants. Most growth parameters were significantly higher in S+ plants. Principal component analysis and hierarchical cluster analysis of 108 metabolites identified by GC–MS clearly demonstrated that in vitro culture had a profound impact on metabolic profile. In vitro S− and S+ plantlets accumulated large quantities of amino acids (specially under S+), photorespiration intermediates, putrescine, tocopherol and organic acids, including oxalic and tartaric acid. However, glycolytic and TCA cycle intermediates were found in lower amount. Under InVR S+ conditions, proline, gamma-aminobutyric acid, sugars and sugar alcohols accumulated in larger amounts. InVR S− plantlets characteristically accumulated large quantity of urea. We suggest that ammonia metabolism was redirected towards urea biosynthesis through urea cycle to sequester nitrogen in condition of low carbon availability. In vitro conditions are causing major disruption in the cellular metabolism, which could produce serious consequences on the capacity of plantlets to adapt to uncontrolled growing conditions and may lead to poor development under these conditions.

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Abbreviations

MS:

Murashige and Skoog (1962) medium

Amax :

Light saturated photosynthesis

S−:

Culture medium without sugar

S+:

Culture medium with sugar

GC–MS:

Gas chromatography–mass spectrometry

TAC:

Tricarboxylic acid cycle

UC:

Urea cycle

GABA:

Gamma-aminobutyric acid

PCA:

Principal component analysis

3PGA:

3 Phosphoglyceric acid

PAs:

Polyamines

Put:

Putrescine

Spd:

Spermidine

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Acknowledgments

Thanks to the Egyptian Higher Education and its Missions General Administration for their financial assistance. The authors also wish to thanks NSERC discovery Grant program for their financial support to Yves Desjardins.

Author information

Correspondence to Ashraf Badr.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Fig.S1

Changes in metabolites extracted from the leaves of potato (Solanum tuberosum L., cv Norland) grown on 3% and 0% sucrose during in vitro rooting and ex vitro acclimatization stages. The results are mean ± SD (n=15), error bars are not shown where they are smaller than the symbol. a, metabolites 1-36; b, metabolites 37-72 ; c, metabolites 73-107 (TIFF 55130 kb)

Fig.S2

Changes in all identified metabolites recovered in methanolic extracts from leaves of potato (Solanum tuberosum L., cv Norland) grown on 3% sucrose, S+ (A, B and C) and 0% sucrose, S- (D, E and F) during in vitro rooting stage (A and D) and ex vitro acclimatization stage (B, C, E and F). Metabolites are shown in order of decreasing normalized peak area in in vitro S+ leaf (A) (TIFF 26721 kb)

Fig.S3

PCA loading plot representing the contribution of individual metabolites to principal component clustering of potato leaves plantlet (Solanum tuberosum L., cv Norland) grown on 0% or 3% sucrose in in vitro rooting stage (TIFF 1259 kb)

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Badr, A., Angers, P. & Desjardins, Y. Comprehensive analysis of in vitro to ex vitro transition of tissue cultured potato plantlets grown with or without sucrose using metabolic profiling technique. Plant Cell Tiss Organ Cult 122, 491–508 (2015). https://doi.org/10.1007/s11240-015-0786-3

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Keywords

  • Potato
  • Micropropagation
  • Stress
  • Gas chromatography–mass spectrometry
  • Principal component analysis
  • Hierarchical cluster analysis
  • Heat map