New Colorimetric Determination of Chloride using Mercuric Thiocyanate and Ferric Ion

Iwasaki, Iwaji; Utsumi, Satori; Ozawa, T.

doi:10.1246/bcsj.25.226

Cited by 246 publications

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“…Sodium and potassium levels were assayed by emission spectrophotometry (Shimazu AA-640). Chloride and betaine contents were determined colorimetrically by the method of Iwasaki et al (15) Figure 1A shows the release of vacuoles under the current method. Lowering the medium osmolarity was essential, but without the addition of CHAPS, more than 20% of the protoplasts retained their cytosolic constituents.…”

Section: Methodsmentioning

confidence: 99%

Sodium, Potassium, Chloride, and Betaine Concentrations in Isolated Vacuoles from Salt-Grown Atriplex gmelini Leaves

Matoh¹,

Watanabe²,

Takáhashi³

1987

Plant Physiol.

116

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Vacuoles were isolated via protoplasts from the leaves of a halophyte Atriplex gmelini C.A.Mey., grown in culture solution supplemented with 250 millimolar NaCl. Lysis of the protoplasts was induced by lowering the medium osmolarity (1.2 to 1.0 molar sorbitol) and adding a detergent, a synthesized cholate derivative, 3-(13-cholamidopropyll dimethylammonio)-l-propanesulfonate at a concentration of 0.5 millimolar and the resulting vacuoles were purified by successive dilution and floatation. Isolated vacuoles contained almost the same concentration of sodium (569 millimolar) and chloride (260 millimolar) as recorded in protoplasts (582 and 254 millimolar, respectively), suggesting that the vacuoles are the major sequestration site of NaCI in leaves of halophytes. Betaine concentration in the protoplasts was about 16 millimolar, while that in vacuoles was only about 0.24 millimolar, indicating that betaine is accumulated in the cytoplasm as a compatible solute.It is generally recognized that the process of osmotic adjustment in dicotyledonous halophytes subjected to saline conditions is achieved primarily by the uptake and accumulation of NaCI into the shoots (7, 24). In the leaves of Atriplex gmelini plants grown in culture solution supplemented with NaCl at 250 mm, the sodium concentration of the tissue amounts to more than 500 mm without any deleterious effects on plant metabolism (21). Since the metabolic reactions occurring in the dicotyledonous halophyte leaves are known to be sensitive to high salt concentration as in the case of the leaves of glycophytes (8, 1 1), it is proposed that the salts responsible for maintaining the leaf solute osmolarity should be sequestered into vacuoles and the difference in the water potential between the cytoplasm and vacuole is compensated by the accumulation of organic solutes such as betaine and proline in the cytoplasm (7, 24). Results of compartmental analysis (32), x-ray microprobe analysis (10, 13), and histochemical analysis (12) have supported this hypothesis. However, the direct determination of the solute concentration in isolated vacuoles from halophytes has not been reported to our knowledge. In order to confirm that vacuoles are the major site of salt sequestration, attempts were made to isolate vacuoles from protoplasts prepared from the leaves of A. gmelini plants ence of 250 mM NaCl during September to December 1985 in a greenhouse were used (21). The lower epidermis was peeled off with a forceps and the leaves were cut into small pieces (0.5 x 1 cm), then floated on an incubation medium. Leaves whose lower epidermis could not be easily peeled off, such as too young and too old leaves, were discarded. The medium consisted of 1.2 M sorbitol, 1 mM Ca(NO3)2, 10 mm ascorbate, 2% Cellulase Onozuka R-10, 1% Macerozyme Onozuka R-10 (Yakult Biochemicals, Nishinomiya, Japan), 0.1% Pectolyase Y-23 (Seishin Pharmaceutical Co. Ltd., Tokyo, Japan), and 20 mM Mes (pH 6.0, adjusted with 2 M Tris solution). Usually a batch of 5 g leaves was incubated in 25 ml of the ...

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Section: Methodsmentioning

confidence: 99%

Sodium, Potassium, Chloride, and Betaine Concentrations in Isolated Vacuoles from Salt-Grown Atriplex gmelini Leaves

Matoh¹,

Watanabe²,

Takáhashi³

1987

Plant Physiol.

116

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“…Na + and K + were determined by flame photometry (GDV Digiflame 2000 with air butane flame). Cl concentration was measured by the mercuric thiocyanate method (Iwasaki et al, 1952).…”

Section: Sample Processingmentioning

confidence: 99%

Acclimation trials of juvenile Italian sturgeon to different salinities: morpho-physiological descriptors

Cataldi¹

1995

Journal of Fish Biology

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In this study the first experimental trials of acclimation to salinities of the Italian sturgeon Acipenser naccarii are described. Preliminary observations regarding some morphological features of the gut, kidney and gills following exposure to 20 and 30‰ salinity are reported, and compared with those displayed by many teleosts in hyper-and hypoosmoregulation. Results are discussed with regard to the osmoregulatory mechanisms and to the physiological limits of adaptability of this species in the second year of its life cycle. Within the Acipenseridae, Acipenser naccarii is generally considered a euryhaline species, but nothing is known about its mechanisms of osmotic homeostasis. This species could be a good candidate for aquaculture in fresh and brackish waters owing to the recent success obtained in artificial reproduction.1995 The Fisheries Society of the British Isles

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“…The levels of Na+ and K+ ions in the supernatant were determined by flame photometry in an atomic absorption spectrophotometer (model 208; Hitachi Ltd., Tokyo, Japan). The C1 ions were quantitated colorimetrically by the method of Iwasaki et al (5). One milliliter of a solution of mercuric thiocyanate and 2 ml of ferric ammonium solution (see below) were added to 1 ml of the supernatant.…”

Section: Methodsmentioning

confidence: 99%

Efflux of sodium ions by a Na+/H+-antiporter during salt stress in the salt-tolerant yeast Zygosaccharomyces rouxii.

Nishi

Yagi

1995

J. Gen. Appl. Microbiol.

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The flux of Nat, Cl-and H+ ions and the accumulation of glycerol in cells of a salt-tolerant yeast, Zygosaccharomyces rouxii, was examined during the initial stages of salt stress. Although the intracellular accumulation of glycerol that was required for osmoregulation began immediately after exposure of the cells to salt stress, rapid influx of Na+ and Cl-ions into cells also occurred. These ions were subsequently extruded from the cells and the intracellular levels of both ions were maintained much lower than those in the external medium. Furthermore, a rapid efflux of protons was induced by activation of the plasma membrane ATPase (PMATPase) immediately after the start of salt stress and then the influx (re-influx) of protons into cells occurred. The addition of a specific inhibitor of PM-ATPase, diethylstilbestrol (DES), and of inhibitors of the energy-generating system, namely sodium azide, antimycin A and dinitrophenol, inhibited the intracellular accumulation of glycerol and activation of PM-ATPase, and they retarded the rapid efflux of protons and the extrusion of Na+ and Cl-ions. The re-influx of protons into cells was inhibited by amphotericin B, probably because of the destruction of the structure of the plasma membrane. In mutants in which rapid activation of PM-ATPase was not observed immediately after the start of salt stress, the efflux of protons was markedly retarded and extrusion of Na+ and C1 ions was also suppressed during salt stress. Amiloride, an inhibitor of the Na+/H+-antiporter, inhibited the re-influx of protons and the efflux of Na+ and Cl-ions, but this inhibitor did not inhibit the accumulation of glycerol, activation of PM-ATPase or the rapid efflux of protons. These results suggest that the Na+/H+-antiporter may play a significant role in extrusion of the ions that enter cells during the initial stages of salt stress.

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New Colorimetric Determination of Chloride using Mercuric Thiocyanate and Ferric Ion

Cited by 246 publications

References 0 publications

Sodium, Potassium, Chloride, and Betaine Concentrations in Isolated Vacuoles from Salt-Grown Atriplex gmelini Leaves

Sodium, Potassium, Chloride, and Betaine Concentrations in Isolated Vacuoles from Salt-Grown Atriplex gmelini Leaves

Acclimation trials of juvenile Italian sturgeon to different salinities: morpho-physiological descriptors

Efflux of sodium ions by a Na+/H+-antiporter during salt stress in the salt-tolerant yeast Zygosaccharomyces rouxii.

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