Use of Per-C-Deuterated <i>myo</i>-Inositol for Study of Cell Wall Synthesis in Germinating Beans

Sasaki, Kohsuke; Nagahashi, Gerald; Gretz, Michael R.; Taylor, Imogen

doi:10.1104/pp.90.2.686

Cited by 8 publications

(3 citation statements)

References 19 publications

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“…Unlike procedures utilizing 2H- (Sasaki et al 1989) or ~3C-labeled sugars (Greve and Labavitch 1991), in which the choice of substrate may influence the distribution of label, sugars were labeled in proportion to their abundance with the 13C02 method.…”

Section: Discussionmentioning

confidence: 99%

Cell wall synthesis during growth and maturation of Nitella internodal cells

Morrison

1993

Planta

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An improved (13)C-density-labeling method was used to study cell wall synthesis in rapidly expanding, slowly expanding and recently mature internodes of Nitella translucens var axillaris (A.Br.) R.D.W. As cells matured, the rate of wall synthesis slowed and the deposition of cellulose microfibrils changed from a predominantly transverse direction in the primary wall of rapidly expanding internodes to a helicoidal array in the secondary wall of mature internodes. The secondary wall was characterized by relatively higher rates of cellulose synthesis and lower rates of pectin synthesis than the primary wall. The synthesis of xyloglucan also decreased markedly at the transition to secondary wall synthesis, while the synthesis of mannose-rich hemicellulose increased. Even though structural differences were striking between the primary and secondary walls of Nitella, compositional differences between the two types of wall were quantitative rather than qualitative.

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Section: Discussionmentioning

confidence: 99%

Cell wall synthesis during growth and maturation of Nitella internodal cells

Morrison

1993

Planta

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“…Detection of the label (and, thus, incorporation into the wall) is accomplished by means of a GC-MS analysis which compares the isotope ratio of selected ion pairs from individual monosaccharide residues. This method is similar to the technique of Sasaki et al (18) who examined the fate of density-labeled inositol in the cell walls of germinating beans but is useful for the analysis of a greater range of monosaccharides because of the more general distribution of carbon from glucose into other sugars. The technique can also be applied to permethylated samples of cell wall polysaccharides and thus provide information regarding incorporation into specific cell wall polymer types.…”

mentioning

confidence: 99%

Cell Wall Metabolism in Ripening Fruit

Lc¹,

Jm²

1991

Plant Physiol.

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A gas chromatographic-mass spectrometric technique utilizing D-[U-13C]glucose as a density label tracer was used to follow the synthesis of cell wall polysaccharides in pericarp discs that were excised from mature green tomato fruit (Lycopersicon esculentum) and allowed to ripen in culture. The biosynthetic capacity of discs from four different maturity stages was examined. Label was differentially incorporated into wall polysaccharides as the discs matured, indicating a change in the nature of wall polymers being synthesized. These differential rates of incorporation are consistent with descriptions of ripening-related cell wall compositional changes previously reported by other authors. Specific changes in wall biosynthesis noted include increased incorporation of xylosyl and mannosyl residues into hemicellulosic cell wall fractions as the discs mature and decreased incorporation of galactosyl residues into chelator-soluble pectins.Digestion of cell wall components has been the primary focus of research for several decades aimed at understanding ripening-related tissue softening of fruits (4,(11)(12)(13)21 saccharides well after the onset of ripening even though the total amount of cell wall material (per gram fruit fresh weight) is steadily decreasing. Synthesis continued into the red maturity stage, and they suggested that these synthetic events might be important in fruit softening. The results of this study generally indicate that cell wall polysaccharide synthesis continued well into the latter stages of tomato fruit ripening, but they provided no clear information concerning the nature of the polymers being synthesized or incorporation into individual monosaccharide residues. We have developed an analytical approach (9) (6). This combination of analytical approach and ripening system allows us to examine in detail incorporation at any stage of the ripening process. Detection of the label (and, thus, incorporation into the wall) is accomplished by means of a GC-MS analysis which compares the isotope ratio of selected ion pairs from individual monosaccharide residues. This method is similar to the technique of Sasaki et al. (18) who examined the fate of density-labeled inositol in the cell walls of germinating beans but is useful for the analysis of a greater range of monosaccharides because of the more general distribution of carbon from glucose into other sugars. The technique can also be applied to permethylated samples of cell wall polysaccharides and thus provide information regarding incorporation into specific cell wall polymer types. In this paper we demonstrate the utility of this density label technique in exploring cell wall synthesis potential and provide data that show that this potential is expressed well into advanced stages of ripening in the tomato fruit. MATERIALS AND METHODS Disc Preparation and TreatmentTomato pericarp discs were prepared from-field grown tomato fruit (Lycopersicon esculentum Mill. cv "Castlemart"), as described in detail by Campbell et al. (6). Briefly, discs (...

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“…Myo-inositol is also considered as a growth enhancer in vitro and may be a carbohydrate source, which are good osmoticum for protoplast isolation (Azad et al 2006). Myo-inositol oxidation pathway has a predominant role in the biosynthesis of new cell walls of the hypocotyl and roots in germinating beans (Sasaki et al 1989). Recently, Abid et al (2009) reported that myo-inositol phosphate synthase (MIPS), which catalyses the first step in inositol biosynthesis are involved in various physiological processes including embryogenesis, seedling growth and resistance to biotic and abiotic stresses.…”

Section: Effect Of Growth Regulators On Cell Division Of Cabbage Protmentioning

confidence: 99%

Myo-inositol increases the plating efficiency of protoplast derived from cotyledon of cabbage (Brassica oleracea var. capitata)

Jie¹,

Kim²,

Jang³

et al. 2011

Journal of Plant Biotechnology

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This study describes the effect of myo-inositol on sustained cell division and plant regeneration from cotyledon-derived protoplast of cabbage (Brassica oleracea var. capitata). Freshly isolated protoplasts were cultured in modified Murashige and Skoog (MS) medium removed ammonia ions and containing 0.4 mg l-1 thiamine･HCl, 100 mg l-1 myo-inositol, 2 mgl-1 2,4-D, 0.5 mgl-1 BA, 30 gl-1 sucrose and several concentrations of myo-inositol (2, 4, 6, 8, 10% (w/v)) as an osmotic stabilizer. After 3 weeks of culture in the dark at 25℃, the plating efficiency of cabbage protoplasts reached to 22.5 ± 2.9% when cultured in modified MS medium supplemented with 2 mgl-1 2,4-D, 0.5 mgl-1 BA, 30 gl-1 sucrose and 8% (w/v) of myo-inositol at a density of 2 x 10 5 protoplasts/ml. Rapidly growing cell colonies after 3 weeks of culture were transferred to the same culture medium removed osmoticum. To induce shoot regeneration from calluses, calluses with about 2 mm in diameter were transferred to the MS medium containing 2 mgl-1 BA and 0.5 mgl-1 NAA. After further three weeks of incubation onto the medium in the light, green shoots were formed on the surface of calluses at a frequency of 30%. Upon transfer to half-strength MS basal medium, roots were formed onto the bottom of regenerated shoots without auxin treatments. These regenerated plantlets were successfully acclimatized to soil transfer, grown to normal mature plants. The cabbage protoplast culture system established in this study could be applied for production of somatic hybrids or cybrids by asymmetric protoplast fusion and mass proliferation of elite somatic clones of cabbage.

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Use of Per-C-Deuterated myo-Inositol for Study of Cell Wall Synthesis in Germinating Beans

Cited by 8 publications

References 19 publications

Cell wall synthesis during growth and maturation of Nitella internodal cells

Cell wall synthesis during growth and maturation of Nitella internodal cells

Cell Wall Metabolism in Ripening Fruit

Myo-inositol increases the plating efficiency of protoplast derived from cotyledon of cabbage (Brassica oleracea var. capitata)

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