Metabolism and development – integration of micro computed tomography data and metabolite profiling reveals metabolic reprogramming from floral initiation to silique development

Abstract

The interrelationship of morphogenesis and metabolism is a poorly studied phenomenon. The main paradigm is that development is controlled by gene expression. The aim of the present study was to correlate metabolism to early and late stages of flower and fruit development in order to provide the basis for the identification of metabolic adjustment and limitations.A highly detailed picture of morphogenesis is achieved using nondestructive micro computed tomography. This technique was used to quantify morphometric parameters of early and late flower development in an Arabidopsis thaliana mutant with synchronized flower initiation. The synchronized flower phenotype made it possible to sample enough early floral tissue otherwise not accessible for metabolomic analysis.The integration of metabolomic and morphometric data enabled the correlation of metabolic signatures with the process of flower morphogenesis. These signatures changed significantly during development, indicating a pronounced metabolic reprogramming in the tissue. Distinct sets of metabolites involved in these processes were identified and were linked to the findings of previous gene expression studies of flower development. High correlations with basic leucine zipper (bZIP) transcription factors and nitrogen metabolism genes involved in the control of metabolic carbon : nitrogen partitioning were revealed.Based on these observations a model for metabolic adjustment during flower development is proposed.

Transcriptomic and lipidomic profiles of glycerolipids during Arabidopsis flower development

“…Dexamethasone (DEX) was applied to the inflorescences of pAP1::AP1‐GR ap1‐1 cal‐5 by dipping in an aqueous solution (pH 7.0) containing 1 μM DEX and 0.015% (v/v) Silwet L‐77. Construction of pAP1::AP1‐GR ap1‐1 cal‐5 was described previously (Bellaire et al ., ).…”

Potato tuber expression of Arabidopsis WRINKLED1 increase triacylglycerol and membrane lipids while affecting central carbohydrate metabolism

“…For the metabolite measurements by GC‐MS analysis, 20 mg of flash‐frozen and lyophilized potato tuber material was ground to a fine powder using a shaking mill and glass beads (5 mm). The polar fraction was extracted and derivatized with 30 μL methoxyamine hydrochloride and 60 μL N‐methyl‐N‐(trimethylsilyl) trifluoroacetamide (MSTFA) as previously described (Bellaire et al ., ) to transform the metabolites into their methoxyimino (MEOX)‐ and trimethylsilyl (TMS)‐ derivatives. allo ‐Inositol was used as an internal standard.…”

“…Error bars, standard deviation (For individual transcript levels used see Table S3). Abbreviations Enzymes: P, plastidial; M, mitochondrial; 2OGDH, 2oxoglutarate dehydrogenase; ACAT, Acetyl-CoA C-acetyltransferase; AGPase, ADP-glucose pyrophosphorylase; CS, citrate synthase; DGAT, diacylglycerol o-acyltransferase; DIT, dicarboxylate transporter; G3P Deh., glycerol-3-phosphate dehydrogenase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GPAT, glycerol-3-phosphate 2-o-acyltransferase; GPT, glucose-6-phosphate phosphate translocator; HMGR, 3-Hydroxy-3-methylglutaryl-CoA reductase; HMGS, hydroxymethylglutaryl-CoA synthase; IDH, isocitrate dehydrogenase; LPAT, 1-Acyl-sn-glycerol- The polar fraction was extracted and derivatized with 30 lL methoxyamine hydrochloride and 60 lL N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) as previously described (Bellaire et al, 2014) to transform the metabolites into their methoxyimino (MEOX)-and trimethylsilyl (TMS)-derivatives. allo-Inositol was used as an internal standard.…”

Identification of the first glyphosate transporter by genomic adaptation

“…Phase separation was induced by adding 250 μl water, samples were centrifuged for 10 min at 1000 g and the polar phase transferred to a new tube. About 200 μl of the polar fraction were dried under a stream of nitrogen and derivatized with 15 μl methoxyamine hydrochloride in pyridine (30 mg/ml) and 30 μl N-methyl-N-(trimethylsilyl) trifluoroacetamide as previously described (Bellaire et al, 2014) to transform metabolites into their methoxyimino-and trimethylsilyl-derivatives. The samples were analysed on an Agilent 5977 N mass selective detector connected to an Agilent 7890B gas chromatograph equipped with a capillary HP5-MS column (30 m × 0.25 mm; 0.25 μm coating thickness; J&W Scientific, Agilent).…”