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In vivo spectroscopy and NMR metabolite fingerprinting approaches to connect the dynamics of photosynthetic and metabolic phenotypes in resurrection plant Haberlea rhodopensis during desiccation and recovery.

Mladenov P, Finazzi G, Bligny R, Moyankova D, Zasheva D, Boisson AM, Brugière S, Krasteva V, Alipieva K, Simova S, Tchorbadjieva M, Goltsev V, Ferro M, Rolland N, Djilianov D - Front Plant Sci (2015)

Bottom Line: The NMR fingerprint shows the significant metabolic changes in several pathways.We found a decline of valine accompanied by lipid degradation during stress, likely to provide alternative carbon sources for sucrose accumulation at late stages of desiccation.This accumulation, as well as the increased levels of glycerophosphodiesters during drought stress could provide osmoprotection to the cells.

View Article: PubMed Central - PubMed

Affiliation: Abiotic Stress Group, Agrobioinstitute, Agricultural Academy Sofia, Bulgaria.

ABSTRACT
The resurrection plant Haberlea rhodopensis was used to study dynamics of drought response of photosynthetic machinery parallel with changes in primary metabolism. A relation between leaf water content and photosynthetic performance was established, enabling us to perform a non-destructive evaluation of the plant water status during stress. Spectroscopic analysis of photosynthesis indicated that, at variance with linear electron flow (LEF) involving photosystem (PS) I and II, cyclic electron flow around PSI remains active till almost full dry state at the expense of the LEF, due to the changed protein organization of photosynthetic apparatus. We suggest that, this activity could have a photoprotective role and prevent a complete drop in adenosine triphosphate (ATP), in the absence of LEF, to fuel specific energy-dependent processes necessary for the survival of the plant, during the late states of desiccation. The NMR fingerprint shows the significant metabolic changes in several pathways. Due to the declining of LEF accompanied by biosynthetic reactions during desiccation, a reduction of the ATP pool during drought was observed, which was fully and quickly recovered after plants rehydration. We found a decline of valine accompanied by lipid degradation during stress, likely to provide alternative carbon sources for sucrose accumulation at late stages of desiccation. This accumulation, as well as the increased levels of glycerophosphodiesters during drought stress could provide osmoprotection to the cells.

No MeSH data available.


Related in: MedlinePlus

Statistical analysis and visualization of changes in metabolomics data of H. rhodopensis during selected states of desiccation and recovery. States of dehydration and recovery are as defined in Figure 1 (A) PCA biplot of the data, contains the measured metabolites identified in H. rhodopensis as loadings represented as numbers, evaluated in samples at various states (as defined in Figure 1) as scores represented with ovals: electric blue for (C) and light blue for R2; violet for D1 and pink for R1; brown for D2 and green for D3. The compounds numbers correspond to assignment of metabolites in NMR spectra (see Figure 3). ∗correspond to metabolites identified with 1H NMR; ∗∗correspond to metabolites identified with 13C NMR; compounds identified with 31P NMR lack asterisks. (B) Hierarchical cluster analysis (HCA)/Heat map clustering and visualization of dynamics of the quantified compounds represented by their log2 transformed averaged meanings, normalized to control levels. P-values for significance of changes during stress treatments were assigned to each metabolite as: n.s.- non significant, ∗- significant, ∗∗-very significant, ∗∗∗- extremely significant.
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Figure 4: Statistical analysis and visualization of changes in metabolomics data of H. rhodopensis during selected states of desiccation and recovery. States of dehydration and recovery are as defined in Figure 1 (A) PCA biplot of the data, contains the measured metabolites identified in H. rhodopensis as loadings represented as numbers, evaluated in samples at various states (as defined in Figure 1) as scores represented with ovals: electric blue for (C) and light blue for R2; violet for D1 and pink for R1; brown for D2 and green for D3. The compounds numbers correspond to assignment of metabolites in NMR spectra (see Figure 3). ∗correspond to metabolites identified with 1H NMR; ∗∗correspond to metabolites identified with 13C NMR; compounds identified with 31P NMR lack asterisks. (B) Hierarchical cluster analysis (HCA)/Heat map clustering and visualization of dynamics of the quantified compounds represented by their log2 transformed averaged meanings, normalized to control levels. P-values for significance of changes during stress treatments were assigned to each metabolite as: n.s.- non significant, ∗- significant, ∗∗-very significant, ∗∗∗- extremely significant.

Mentions: Combining 13C, 31P, and 1H NMR, we were able to identify and quantify a total of 36 metabolites during dehydration and recovery in H. rhodopensis (Figure 3, Supplementary Table S3). Almost all of the compounds represented in H. rhodopensis showed significant changes during stress (Supplementary Table S3). Sucrose, valine, fructose, glucose, citrate, ethanol, and gucose-6-P were with the highest amounts in the extracts (Supplementary Table S3). The performed PCA explained 70% of the total variance of metabolites within the samples by the two main components. The first group of metabolites (loadings) with highest contribution in PC1 is: phosphoenolpyruvate (PEP), glucose, dihydroxyacetone phosphate (DHAP), glucose 6-phosphate (G6P) and ATP (Figure 4A, loading numbers: 18, 4∗∗, 1, 3, 19) as positive ones and the second group of main negatives are: glycerophosphocholine (GPC), sucrose and glycerophosphoethanolamine (GPE), (Figure 4A, loading numbers: 17, 3∗, 15). Therefore, the leaf samples (scores), in C and R2 state, grouped in the negative scale, are separated from the others due to their highest content of glycolytic intermediates and ATP and low levels of sucrose, GPC and GPE (Figure 4A, scores) on the contrast to the fully dry leaf samples which are extremely in positive scale (Figure 4A, scores). The PC2 separate mainly D3 state in the positive scale and D1 and D2 in the negative from the other samples (Figure 4A, scores), according to the higher content of NADP, ADP, glycerophosphoglycerol (GPG) and glycerol 3-phosphate (G3P) in D3 (Figure 4A, loading numbers: 22, 20, 13, 6), and the higher content of ethanolamine and gamma-aminobutyric acid (GABA) in D1 and D2, respectively (Figure 4A, loading number: 5∗, 6∗). The HCA/Heat map (Figure 4B) showed the same group pattern of the treatments and metabolite changes as PCA, but provided more detailed visualization of the metabolite changes during stress. The metabolites of the first main cluster – bis(glycerophospho)glycerol (GPGP), mannose 6-phosphate (M6P), ATP, and PEP showed a drastic decrease from D1–D3 stages (Figure 4B, rows, top cluster). G6P, DHAP, citrate, valine, glucose, fructose 6-phosphate (F6P), glyceraldehyde phosphate (GAP), phosphatidylethanolamine (PE), decreased immediately after the onset of water deficit and are grouped in the second main cluster (Figure 4B, rows, red group). Phosphocholine (P-choline) is separated from this cluster according to the lower levels in R2. The third main cluster is presented by two different groups. The first one (blue) – ADP, NADP, UDP-glucose (UDP-glu), GPG, G3P, and 6-phosphogluconate groups metabolites with more or less decreased concentrations in D1 and D2 states, with a tendency of accumulation during D2–D3 state transition. The second group (light blue) combines metabolites with relatively stable concentrations during drought stress – 3-phosphoglycerate (PGA), fructose, ethanol, glutamate, aspartate, alanine, and fumarate. The metabolites with increased concentration during drought stress represent the fourth main cluster (Figure 4B rows, bottom cluster). GPC, GPE, sucrose (contributing for the separation of D3 in PC1 positive scale) are grouped in pink. The second group (green) of this cluster combines metabolites with heterogeneity in their accumulation during drought. NADPH, glycerophosphoinositol (GPI) and AMP increased from D2–D3. Succinate, choline, ethanolamine, and GABA accumulated during D1 and D2 states and decreased in D3 state, thus explaining the separation of these states in PC2. After rewatering (R1), the metabolites recovered their abundance to levels close to D1 state. At R2 state, the contents of the metabolites were more or less close to control (C) state.


In vivo spectroscopy and NMR metabolite fingerprinting approaches to connect the dynamics of photosynthetic and metabolic phenotypes in resurrection plant Haberlea rhodopensis during desiccation and recovery.

Mladenov P, Finazzi G, Bligny R, Moyankova D, Zasheva D, Boisson AM, Brugière S, Krasteva V, Alipieva K, Simova S, Tchorbadjieva M, Goltsev V, Ferro M, Rolland N, Djilianov D - Front Plant Sci (2015)

Statistical analysis and visualization of changes in metabolomics data of H. rhodopensis during selected states of desiccation and recovery. States of dehydration and recovery are as defined in Figure 1 (A) PCA biplot of the data, contains the measured metabolites identified in H. rhodopensis as loadings represented as numbers, evaluated in samples at various states (as defined in Figure 1) as scores represented with ovals: electric blue for (C) and light blue for R2; violet for D1 and pink for R1; brown for D2 and green for D3. The compounds numbers correspond to assignment of metabolites in NMR spectra (see Figure 3). ∗correspond to metabolites identified with 1H NMR; ∗∗correspond to metabolites identified with 13C NMR; compounds identified with 31P NMR lack asterisks. (B) Hierarchical cluster analysis (HCA)/Heat map clustering and visualization of dynamics of the quantified compounds represented by their log2 transformed averaged meanings, normalized to control levels. P-values for significance of changes during stress treatments were assigned to each metabolite as: n.s.- non significant, ∗- significant, ∗∗-very significant, ∗∗∗- extremely significant.
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Related In: Results  -  Collection

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Figure 4: Statistical analysis and visualization of changes in metabolomics data of H. rhodopensis during selected states of desiccation and recovery. States of dehydration and recovery are as defined in Figure 1 (A) PCA biplot of the data, contains the measured metabolites identified in H. rhodopensis as loadings represented as numbers, evaluated in samples at various states (as defined in Figure 1) as scores represented with ovals: electric blue for (C) and light blue for R2; violet for D1 and pink for R1; brown for D2 and green for D3. The compounds numbers correspond to assignment of metabolites in NMR spectra (see Figure 3). ∗correspond to metabolites identified with 1H NMR; ∗∗correspond to metabolites identified with 13C NMR; compounds identified with 31P NMR lack asterisks. (B) Hierarchical cluster analysis (HCA)/Heat map clustering and visualization of dynamics of the quantified compounds represented by their log2 transformed averaged meanings, normalized to control levels. P-values for significance of changes during stress treatments were assigned to each metabolite as: n.s.- non significant, ∗- significant, ∗∗-very significant, ∗∗∗- extremely significant.
Mentions: Combining 13C, 31P, and 1H NMR, we were able to identify and quantify a total of 36 metabolites during dehydration and recovery in H. rhodopensis (Figure 3, Supplementary Table S3). Almost all of the compounds represented in H. rhodopensis showed significant changes during stress (Supplementary Table S3). Sucrose, valine, fructose, glucose, citrate, ethanol, and gucose-6-P were with the highest amounts in the extracts (Supplementary Table S3). The performed PCA explained 70% of the total variance of metabolites within the samples by the two main components. The first group of metabolites (loadings) with highest contribution in PC1 is: phosphoenolpyruvate (PEP), glucose, dihydroxyacetone phosphate (DHAP), glucose 6-phosphate (G6P) and ATP (Figure 4A, loading numbers: 18, 4∗∗, 1, 3, 19) as positive ones and the second group of main negatives are: glycerophosphocholine (GPC), sucrose and glycerophosphoethanolamine (GPE), (Figure 4A, loading numbers: 17, 3∗, 15). Therefore, the leaf samples (scores), in C and R2 state, grouped in the negative scale, are separated from the others due to their highest content of glycolytic intermediates and ATP and low levels of sucrose, GPC and GPE (Figure 4A, scores) on the contrast to the fully dry leaf samples which are extremely in positive scale (Figure 4A, scores). The PC2 separate mainly D3 state in the positive scale and D1 and D2 in the negative from the other samples (Figure 4A, scores), according to the higher content of NADP, ADP, glycerophosphoglycerol (GPG) and glycerol 3-phosphate (G3P) in D3 (Figure 4A, loading numbers: 22, 20, 13, 6), and the higher content of ethanolamine and gamma-aminobutyric acid (GABA) in D1 and D2, respectively (Figure 4A, loading number: 5∗, 6∗). The HCA/Heat map (Figure 4B) showed the same group pattern of the treatments and metabolite changes as PCA, but provided more detailed visualization of the metabolite changes during stress. The metabolites of the first main cluster – bis(glycerophospho)glycerol (GPGP), mannose 6-phosphate (M6P), ATP, and PEP showed a drastic decrease from D1–D3 stages (Figure 4B, rows, top cluster). G6P, DHAP, citrate, valine, glucose, fructose 6-phosphate (F6P), glyceraldehyde phosphate (GAP), phosphatidylethanolamine (PE), decreased immediately after the onset of water deficit and are grouped in the second main cluster (Figure 4B, rows, red group). Phosphocholine (P-choline) is separated from this cluster according to the lower levels in R2. The third main cluster is presented by two different groups. The first one (blue) – ADP, NADP, UDP-glucose (UDP-glu), GPG, G3P, and 6-phosphogluconate groups metabolites with more or less decreased concentrations in D1 and D2 states, with a tendency of accumulation during D2–D3 state transition. The second group (light blue) combines metabolites with relatively stable concentrations during drought stress – 3-phosphoglycerate (PGA), fructose, ethanol, glutamate, aspartate, alanine, and fumarate. The metabolites with increased concentration during drought stress represent the fourth main cluster (Figure 4B rows, bottom cluster). GPC, GPE, sucrose (contributing for the separation of D3 in PC1 positive scale) are grouped in pink. The second group (green) of this cluster combines metabolites with heterogeneity in their accumulation during drought. NADPH, glycerophosphoinositol (GPI) and AMP increased from D2–D3. Succinate, choline, ethanolamine, and GABA accumulated during D1 and D2 states and decreased in D3 state, thus explaining the separation of these states in PC2. After rewatering (R1), the metabolites recovered their abundance to levels close to D1 state. At R2 state, the contents of the metabolites were more or less close to control (C) state.

Bottom Line: The NMR fingerprint shows the significant metabolic changes in several pathways.We found a decline of valine accompanied by lipid degradation during stress, likely to provide alternative carbon sources for sucrose accumulation at late stages of desiccation.This accumulation, as well as the increased levels of glycerophosphodiesters during drought stress could provide osmoprotection to the cells.

View Article: PubMed Central - PubMed

Affiliation: Abiotic Stress Group, Agrobioinstitute, Agricultural Academy Sofia, Bulgaria.

ABSTRACT
The resurrection plant Haberlea rhodopensis was used to study dynamics of drought response of photosynthetic machinery parallel with changes in primary metabolism. A relation between leaf water content and photosynthetic performance was established, enabling us to perform a non-destructive evaluation of the plant water status during stress. Spectroscopic analysis of photosynthesis indicated that, at variance with linear electron flow (LEF) involving photosystem (PS) I and II, cyclic electron flow around PSI remains active till almost full dry state at the expense of the LEF, due to the changed protein organization of photosynthetic apparatus. We suggest that, this activity could have a photoprotective role and prevent a complete drop in adenosine triphosphate (ATP), in the absence of LEF, to fuel specific energy-dependent processes necessary for the survival of the plant, during the late states of desiccation. The NMR fingerprint shows the significant metabolic changes in several pathways. Due to the declining of LEF accompanied by biosynthetic reactions during desiccation, a reduction of the ATP pool during drought was observed, which was fully and quickly recovered after plants rehydration. We found a decline of valine accompanied by lipid degradation during stress, likely to provide alternative carbon sources for sucrose accumulation at late stages of desiccation. This accumulation, as well as the increased levels of glycerophosphodiesters during drought stress could provide osmoprotection to the cells.

No MeSH data available.


Related in: MedlinePlus