On the use of 31P NMR for the quantification of hydrosoluble phosphorus-containing compounds in coral host tissues and cultured zooxanthellae

Abstract

31P Nuclear Magnetic Resonance (NMR) was assessed to investigate the phosphorus-containing compounds present in the tissues of the scleractinian coral Stylophora pistillata as well as of cultured zooxanthellae (CZ). Results showed that phosphorus-containing compounds observed in CZ were mainly phosphate and phosphate esters. Phosphate accounted for 19 ± 2% of the total phosphorus compounds observed in CZ maintained under low P-levels (0.02 μM). Adding 5 mM of dissolved inorganic phosphorus (KH2PO4) to the CZ culture medium led to a 3.1-fold increase in intracellular phosphate, while adding 5 mM of dissolved organic phosphorus led to a reduction in the concentration of phosphorus compounds, including a 2.5-fold intracellular phosphate decrease. In sharp contrast to zooxanthellae, the host mainly contained phosphonates, and to a lesser extent, phosphate esters and phosphate. Two-months of host starvation decreased the phosphate content by 2.4 fold, while bleaching of fed corals did not modify this content. Based on 31P NMR analyses, this study highlights the importance of phosphonates in the composition of coral host tissues, and illustrates the impact of phosphorus availability on the phosphorus composition of host tissues and CZ, both through feeding of the host and inorganic phosphorus enrichment of the CZ.

Combined computational and experimental study about the incorporation of phosphorus into the structure of graphene oxide

“…In fact, the existence of this type of bonding is energetically viable, as detailed in the discussion of the data in Table 3, considering that the synthesis conditions used in this work (with concentrated H 3 PO 4 and H 2 SO 4 solutions at a temperature of 50 °C) can lead to the formation of phosphate derivatives containing P–C bonds according to reactions such as the one indicated in Scheme 1. 47,85,87 Therefore, the possibility of the occurrence of both P–O–C (mainly) and P–C bonds in the P/GO hybrids here described is well supported by the experimental and computational results presented in this work.…”

“…95,96 The calculated value also agrees well with chemical shifts reported in the literature (around 0 ppm) for concentrated H 3 PO 4 solutions and solid materials (such as activated carbons) containing adsorbed H 3 PO 4 . 35,78,85,94 As for the 13 C nuclei in the graphene layer, the calculated chemical shifts fall in the range expected for signals due to carbon atoms in sp 2 -hybridized regions (126–128 ppm). 9,35,49,52,81,82 This shows that the simple adsorption of a H 3 PO 4 molecule does not cause detectable changes in the chemical shifts of the 13 C nuclei in the graphene layer.…”

Combined computational and experimental study about the incorporation of phosphorus into the structure of graphene oxide

“…In fact, the existence of this type of bonding is energetically viable, as detailed in the discussion of the data in Table 3, considering that the synthesis conditions used in this work (with concentrated H 3 PO 4 and H 2 SO 4 solutions at a temperature of 50 °C) can lead to the formation of phosphate derivatives containing P–C bonds according to reactions such as the one indicated in Scheme 1. 47,85,87 Therefore, the possibility of the occurrence of both P–O–C (mainly) and P–C bonds in the P/GO hybrids here described is well supported by the experimental and computational results presented in this work.…”

“…95,96 The calculated value also agrees well with chemical shifts reported in the literature (around 0 ppm) for concentrated H 3 PO 4 solutions and solid materials (such as activated carbons) containing adsorbed H 3 PO 4 . 35,78,85,94 As for the 13 C nuclei in the graphene layer, the calculated chemical shifts fall in the range expected for signals due to carbon atoms in sp 2 -hybridized regions (126–128 ppm). 9,35,49,52,81,82 This shows that the simple adsorption of a H 3 PO 4 molecule does not cause detectable changes in the chemical shifts of the 13 C nuclei in the graphene layer.…”

Phosphorus metabolism of reef organisms with algal symbionts

“…The concentration of free phosphates in symbionts ranges between 1.3 and 10.3 fmol P.symbiont -1 or 5.0-38.3 mmol P.L -1 (Jackson 1989), which is in agreement with concentrations measured in phytoplankton cells (between 1 and 15 mmol P.L -1 ) (Raven 1974, Reed et al 1981. Intracellular phosphate concentrations tend to increase with cell age, or in response to P enrichment of seawater (Godinot et al 2016), as previously demonstrated also for phytoplankton (Watanabe et al 1987, Feuillade et al 1995. Moreover, both host tissue of anthozoans and their dinoflagellate symbionts contain several types of phosphonates (Kittredge and Roberts 1969, Steen 1986, Rands et al 1992, Godinot et al 2016), which represent a major pool of phosphorus for marine microorganisms in general (Martinez et al 2010, Villarreal-Chiu et al 2012, and cnidarians in particular (Quin 1964, Henderson et al 1972, Steen 1986, accounting for between 10-50% of cellular particulate phosphorus.…”

“…Intracellular phosphate concentrations tend to increase with cell age, or in response to P enrichment of seawater (Godinot et al. ), as previously demonstrated also for phytoplankton (Watanabe et al. , Feuillade et al.…”

Exploiting the Versatility of Electrosynthesized Amino-bis(methylene phosphonate) Synthons via Coupling: Enriching the Phosphonate Linker “Toolbox” Toward Metal-Phosphonate Coordination Networks

“…In any case, no phosphonate ester signal was detected in the 31 P NMR spectra of any of the products, since those appear in a very different chemical shift range (−22 to −24 ppm) compared to the ones observed (5−14 ppm). 39,40 Even in the highly alkaline environment, in which the phosphonate groups have a high anionic charge and one would expect them to act as nucleophiles, no significant formation of phosphonate esters was observed in our experiments.…”

Combined computational and experimental study about the incorporation of phosphorus into the structure of graphene oxide

“…In fact, the existence of this type of bonding is energetically viable, as detailed in the discussion of the data in Table 3, considering that the synthesis conditions used in this work (with concentrated H 3 PO 4 and H 2 SO 4 solutions at a temperature of 50 °C) can lead to the formation of phosphate derivatives containing P–C bonds according to reactions such as the one indicated in Scheme 1. 47,85,87 Therefore, the possibility of the occurrence of both P–O–C (mainly) and P–C bonds in the P/GO hybrids here described is well supported by the experimental and computational results presented in this work.…”

“…95,96 The calculated value also agrees well with chemical shifts reported in the literature (around 0 ppm) for concentrated H 3 PO 4 solutions and solid materials (such as activated carbons) containing adsorbed H 3 PO 4 . 35,78,85,94 As for the 13 C nuclei in the graphene layer, the calculated chemical shifts fall in the range expected for signals due to carbon atoms in sp 2 -hybridized regions (126–128 ppm). 9,35,49,52,81,82 This shows that the simple adsorption of a H 3 PO 4 molecule does not cause detectable changes in the chemical shifts of the 13 C nuclei in the graphene layer.…”

Phosphorus metabolism of reef organisms with algal symbionts

“…The concentration of free phosphates in symbionts ranges between 1.3 and 10.3 fmol P.symbiont -1 or 5.0-38.3 mmol P.L -1 (Jackson 1989), which is in agreement with concentrations measured in phytoplankton cells (between 1 and 15 mmol P.L -1 ) (Raven 1974, Reed et al 1981. Intracellular phosphate concentrations tend to increase with cell age, or in response to P enrichment of seawater (Godinot et al 2016), as previously demonstrated also for phytoplankton (Watanabe et al 1987, Feuillade et al 1995. Moreover, both host tissue of anthozoans and their dinoflagellate symbionts contain several types of phosphonates (Kittredge and Roberts 1969, Steen 1986, Rands et al 1992, Godinot et al 2016), which represent a major pool of phosphorus for marine microorganisms in general (Martinez et al 2010, Villarreal-Chiu et al 2012, and cnidarians in particular (Quin 1964, Henderson et al 1972, Steen 1986, accounting for between 10-50% of cellular particulate phosphorus.…”

“…Intracellular phosphate concentrations tend to increase with cell age, or in response to P enrichment of seawater (Godinot et al. ), as previously demonstrated also for phytoplankton (Watanabe et al. , Feuillade et al.…”

Exploiting the Versatility of Electrosynthesized Amino-bis(methylene phosphonate) Synthons via Coupling: Enriching the Phosphonate Linker “Toolbox” Toward Metal-Phosphonate Coordination Networks

“…In any case, no phosphonate ester signal was detected in the 31 P NMR spectra of any of the products, since those appear in a very different chemical shift range (−22 to −24 ppm) compared to the ones observed (5−14 ppm). 39,40 Even in the highly alkaline environment, in which the phosphonate groups have a high anionic charge and one would expect them to act as nucleophiles, no significant formation of phosphonate esters was observed in our experiments.…”

Combined computational and experimental study about the incorporation of phosphorus into the structure of graphene oxide

“…In fact, the existence of this type of bonding is energetically viable, as detailed in the discussion of the data in Table 3, considering that the synthesis conditions used in this work (with concentrated H 3 PO 4 and H 2 SO 4 solutions at a temperature of 50 °C) can lead to the formation of phosphate derivatives containing P–C bonds according to reactions such as the one indicated in Scheme 1. 47,85,87 Therefore, the possibility of the occurrence of both P–O–C (mainly) and P–C bonds in the P/GO hybrids here described is well supported by the experimental and computational results presented in this work.…”

“…95,96 The calculated value also agrees well with chemical shifts reported in the literature (around 0 ppm) for concentrated H 3 PO 4 solutions and solid materials (such as activated carbons) containing adsorbed H 3 PO 4 . 35,78,85,94 As for the 13 C nuclei in the graphene layer, the calculated chemical shifts fall in the range expected for signals due to carbon atoms in sp 2 -hybridized regions (126–128 ppm). 9,35,49,52,81,82 This shows that the simple adsorption of a H 3 PO 4 molecule does not cause detectable changes in the chemical shifts of the 13 C nuclei in the graphene layer.…”

Phosphorus metabolism of reef organisms with algal symbionts

“…The concentration of free phosphates in symbionts ranges between 1.3 and 10.3 fmol P.symbiont -1 or 5.0-38.3 mmol P.L -1 (Jackson 1989), which is in agreement with concentrations measured in phytoplankton cells (between 1 and 15 mmol P.L -1 ) (Raven 1974, Reed et al 1981. Intracellular phosphate concentrations tend to increase with cell age, or in response to P enrichment of seawater (Godinot et al 2016), as previously demonstrated also for phytoplankton (Watanabe et al 1987, Feuillade et al 1995. Moreover, both host tissue of anthozoans and their dinoflagellate symbionts contain several types of phosphonates (Kittredge and Roberts 1969, Steen 1986, Rands et al 1992, Godinot et al 2016), which represent a major pool of phosphorus for marine microorganisms in general (Martinez et al 2010, Villarreal-Chiu et al 2012, and cnidarians in particular (Quin 1964, Henderson et al 1972, Steen 1986, accounting for between 10-50% of cellular particulate phosphorus.…”

“…Intracellular phosphate concentrations tend to increase with cell age, or in response to P enrichment of seawater (Godinot et al. ), as previously demonstrated also for phytoplankton (Watanabe et al. , Feuillade et al.…”

Exploiting the Versatility of Electrosynthesized Amino-bis(methylene phosphonate) Synthons via Coupling: Enriching the Phosphonate Linker “Toolbox” Toward Metal-Phosphonate Coordination Networks

“…In any case, no phosphonate ester signal was detected in the 31 P NMR spectra of any of the products, since those appear in a very different chemical shift range (−22 to −24 ppm) compared to the ones observed (5−14 ppm). 39,40 Even in the highly alkaline environment, in which the phosphonate groups have a high anionic charge and one would expect them to act as nucleophiles, no significant formation of phosphonate esters was observed in our experiments.…”