Metabolism control over plant growth and development: A case for trehalose metabolism.

Principle investigator: Henriette Schluepmann

Collaborations at UU: Javier Sastre-Torano, Ad de Jong and Govert Somsen (Dept. Pharmacie); Lidija Berke and Gabino Sanchez-Perez (Dept. Biology).

International Collaborations: Matthew Paul (Rothamstead Research, UK); Astrid Wingler (University College London, UK), Peter Geigenberger and Mark Stitt (MPI-Golm, Germany), Anika Wiese-Klinkenberg (Forschungszentrum Juelich, Germany); Youichi Kondou and Minami Mitsui (RIKEN-Yokohama, Japan).

Students in 2011: Dr. Prapti Sedijani, Thibault Krommenhoeck, Niek Schrier and Max Wellenstein.

The precursor of trehalose biosynthesis, trehalose-6-phosphate (T6P), is essential for development and controls carbon utilization in Arabidospis thaliana seedlings. Furthermore, T6P accumulation in the absence of available carbon causes growth arrest when Arabidopsis seedlings are supplied with 100 mM trehalose. But what are the mechanisms involved?

In 2008, we had been able to establish a medium throughput method to determine T6P levels in plant extracts that has since evolved. The method combined liquid and solid-phase extractions before separation and detection using HPLC/MS, and more recently CE/MS. In 2011 we have improved the method to achieve high throughput HILIC-UPLC/MS detection of phosphorylated intermediates of metabolism.

Through collaboration with Dr. Matthew Paul at Rothamstead Research (UK) we report T6P inhibition of SnRK1 kinase activity showing for the first time that T6P controls growth and carbon utilization by way of a central kinase signaling network in 2009. The collaboration further uncovered a remarkable accumulation of T6P during wheat embryo development in 2011.

Feeding trehalose to Arabidopsis seedlings causes a reversal of carbon allocation with starch accumulating in cotyledons and no starch and growth arrest at the apical meristems (Figure 1). To understand this phenomenon we are using a genetic approach. We have isolated and characterized mutants that overcome the growth inhibition on 100 mM trehalose. A subset of these mutants is affected in genes known to be otherwise involved in nutrient stress responses and these include mutants with altered SnRK1 activity and bZIP11 expression: we conclude that on trehalose medium, T6P accumulation inhibits SnRK1 activity and thence growth in Arabidopsis. Through a collaboration with Dr. Astrid Wingler at University College London (UK) we studied the role of T6P in determining the onset of senescence. Work in progress is aimed at understanding what links trehalose metabolism to carbon allocation and developmental responses.

Figure 1 Feeding trehalose to Arabidopsis seedlings causes reversal of carbon allocation. Seedlings were grown on 100 mM trehalose for 14 days, destained, then starch stained using Lugol. wt, wild type seedlings accumulate large quantities of starch in the cotyledons, the apical meristems are growth arrested and no starch is seen in the columella of the roots. pgm1, mutants of the chloroplastic phosphoglucomutase lack the precursor of starch synthesis and so are unable to make starch yet also stop growing on trehalose. Reversal of carbon allocation by trehalose is therefore not caused by induced starch accumulation. 

A new research line is now emerging as we attempt to understand the tremendous growth rates achieved by the floating fern Azolla and test this fern’s applications for biomass cultivation in a consortium with Gert-Jan Reichert and Peter Bijl (Geo faculty at UU), Ellen van Donk and Liesbeth Bakker (UU and NIOO), Adrie van der Werf (Wageningen University) and Klaas Timmermans and Henk Brinkhuis (NIOZ).

Student projects are available on the Azolla research as well as on the more fundamental research of metabolite control over growth.

We remain open for collaborations and new lines of research.

Figure 2 The fern Azolla/cyanobacteria symbiosis, one of the fastest growing plants on earth.

Publications:

Wingler A, Delatte TL, O'Hara LE, Primavesi LF, Jhurreea D, Paul MJ, Schluepmann H. (2012) Trehalose 6-phosphate is required for the onset of leaf senescence associated with high carbon availability. Plant Physiol. 158(3):1241-51. Epub 2012 Jan 13.

Sastre Toraño J, Delatte TL, Schluepmann H, Smeekens SC, de Jong GJ, Somsen GW. (2012) Determination of trehalose-6-phosphate in Arabidopsis thaliana seedlings by hydrophilic-interaction liquid chromatography-mass spectrometry. Anal Bioanal Chem. Epub 2012 Mar 27.

Schluepmann H, Berke L, Sanchez-Perez GF. (2011) Metabolism control over growth: a case for trehalose-6-phosphate in plants. J Exp Bot. Epub 2011 Nov 4.

Delatte TL, Sedijani P, Kondou Y, Matsui M, de Jong GJ, Somsen GW, Wiese-Klinkenberg A, Primavesi LF, Paul MJ, Schluepmann H. (2011) Growth arrest by trehalose-6-phosphate: an astonishing case of primary metabolite control over growth by way of the SnRK1 signaling pathway. Plant Physiol. 157(1):160-74. Epub 2011 Jul 13.

Martínez-Barajas E, Delatte T, Schluepmann H, de Jong GJ, Somsen GW, Nunes C, Primavesi LF, Coello P, Mitchell RA, Paul MJ. (2011) Wheat grain development is characterized by remarkable trehalose 6-phosphate accumulation pregrain filling: tissue distribution and relationship to SNF1-related protein kinase1 activity. Plant Physiol. 156(1):373-81.

Ma J, Hanssen M, Lundgren K, Hernández L, Delatte T, Ehlert A, Liu CM, Schluepmann H, Dröge-Laser W, Moritz T, Smeekens S, Hanson J. (2011) The sucrose-regulated Arabidopsis transcription factor bZIP11 reprograms metabolism and regulates trehalose metabolism. New Phytol. 191(3):733-45.

Delatte TL, Schluepmann H, Smeekens SC, de Jong GJ, Somsen GW. (2011) Capillary electrophoresis-mass spectrometry analysis of trehalose-6-phosphate in Arabidopsis thaliana seedlings. Anal Bioanal Chem. 400(4):1137-44.

Paul MJ, Jhurreea D, Zhang Y, Primavesi LF, Delatte T, Schluepmann H, Wingler A. (2010) Upregulation of biosynthetic processes associated with growth by trehalose 6-phosphate. Plant Signal Behav. 2010 Apr;5(4):386-92. Epub 2010 Apr 25.

Schluepmann, H & Paul, M (2009) The Arabidopsis Book. Rockville, MD: American Society of Plant Biologists. doi: 10. 1199/tab. 0122 http://www/.aspb. org/publications/arabidopsis/.

Delatte T L, Selman MH, Schluepmann H, Somsen GW, Smeekens, S C & de Jong, G J. (2009) Determination of trehalose-6-phosphate in Arabidopsis seedlings by successive extractions followed by anion exchange chromatography-mass spectrometry. Anal. Biochem. 389, 12-17.

Zhang Y, Primavesi L F, Jhurreea D, Andralojc P J, Mitchell R A, Powers S J, Schluepmann H, Delatte T, Wingler A, Paul M J. (2009) Plant Physiol 149, 1860-1871.

Kolbe A, Tiessen A, Schluepmann H, Paul M, Ulrich S, Geigenberger P. (2005) Trehalose 6-phosphate regulates starch synthesis via posttranslational redox activation of ADP-glucose pyrophosphorylase. Proc Natl Acad Sci U S A. 2005 Aug 2;102(31):11118-23.

Schluepmann H, van Dijken A, Aghdasi M, Wobbes B, Paul M, Smeekens S (2004) Trehalose mediated growth inhibition of Arabidopsis seedlings is due to trehalose-6-phosphate accumulation. Plant Physiol. 135(2):879-90.

Pellny TK, Ghannoum O, Conroy JP, Schluepmann H, Smeekens S, Andralojc J, Krause KP, Goddijn O, Paul MJ. (2004) Genetic modification of photosynthesis with E. coli genes for trehalose synthesis. Plant Biotechnol J. 2(1):71-82.

van Dijken AJ, Schluepmann H, Smeekens SC. (2004) Arabidopsis trehalose-6-phosphate synthase 1 is essential for normal vegetative growth and transition to flowering. Plant Physiol. 135(2):969-77.

Schluepmann H, Pellny T, van Dijken A, Smeekens S, Paul M. (2003) Trehalose 6-phosphate is indispensable for carbohydrate utilization and growth in Arabidopsis thaliana. Proc Natl Acad Sci U S A. 100(11):6849-54.

Schlupmann H, Bacic A, Read SM. (1994) Uridine Diphosphate Glucose Metabolism and Callose Synthesis in Cultured Pollen Tubes of Nicotiana alata Link et Otto. Plant Physiol. 105(2):659-670.

Schlüpmann H, Bacic A, Read SM. (1993) A novel callose synthase from pollen tubes of Nicotiana. Planta 191: 470–481

Peterhans A, Schlüpmann H, Basse C, Paszkowski J. (1990) Intrachromosomal recombination in plants. EMBO J. 9(11):3437-45.