Pectate distribution and esterification in Dubautia leaves and soybean nodules, studied with a fluorescent hybridization probe

Valerie Vreeland, Suzanne R. Morse, Robert H Robichaux, Kathleen L. Miller, Sui Sheng T Hua, Watson M. Laetsch

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Carbohydrate-hybridization probes (Vreeland and Laetsch, 1989, Planta (177, 423-434) were used to localize the homogalacturonan (pectate) component of pectins in the cell walls of leaves and soybean root nodules. Leaves of two species of the dicotyledon Dubautia were compared; these species contain much pectin but differ in their tissue water relations with respect to their cell-wall properties. Maturation of the primary cell walls in nodules was studied in the Bradyrhizobium japonicum-Glycine max symbiosis. Probe labelling was based on the divalent-cation-mediated association between pectate in tissue sections and fluorescein-conjugated pectate fragments. Pectate was also labelled by mixed-dimer formation with fluorescent polyguluronate derived from alginate. The specificity of the probe for unesterified polygalacturonate was indicated by increased cell-wall labelling after chemical or enzymatic deesterification of tissue sections, in contrast to elimination of labelling by chemical esterification. Postfixation of tissue sections improved retention of soluble pectate. Pectate differences were found in the leaves among cell types, in degree of esterification, and between plant species. The cell walls of soybean nodules were strongly labelled by the pectate probe in nodules one week and three weeks after infection. Pectate was more highly esterified in the central infected zone than in the surrouding cortex. Within the infected zone, walls of uninfected cells and infected cells were similarly labelled by the pectate probe. The results indicate that the pectate molecular probe provides detailed information on pectate distribution at the cellular level for investigations of cell-wall structure, development and physiology.

Original languageEnglish (US)
Pages (from-to)435-446
Number of pages12
JournalPlanta
Volume177
Issue number4
DOIs
StatePublished - Apr 1989
Externally publishedYes

Fingerprint

Dubautia
Esterification
esterification
Fluorescent Dyes
Soybeans
cell walls
soybeans
probes (equipment)
Cell Wall
leaves
pectins
Bradyrhizobium japonicum
root nodules
alginates
fluorescein
Plantae
hybridization probes
polygalacturonic acid
Magnoliopsida
symbiosis

Keywords

  • Bradyrhizobium
  • Cell wall (pectate distribution)
  • Dubautia
  • Glycine (root-nodule cell wall)
  • Hybridization probe
  • Leaf (cell wall)
  • Pectate
  • Root nodule (cell wall)

ASJC Scopus subject areas

  • Plant Science

Cite this

Pectate distribution and esterification in Dubautia leaves and soybean nodules, studied with a fluorescent hybridization probe. / Vreeland, Valerie; Morse, Suzanne R.; Robichaux, Robert H; Miller, Kathleen L.; Hua, Sui Sheng T; Laetsch, Watson M.

In: Planta, Vol. 177, No. 4, 04.1989, p. 435-446.

Research output: Contribution to journalArticle

Vreeland, Valerie ; Morse, Suzanne R. ; Robichaux, Robert H ; Miller, Kathleen L. ; Hua, Sui Sheng T ; Laetsch, Watson M. / Pectate distribution and esterification in Dubautia leaves and soybean nodules, studied with a fluorescent hybridization probe. In: Planta. 1989 ; Vol. 177, No. 4. pp. 435-446.
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abstract = "Carbohydrate-hybridization probes (Vreeland and Laetsch, 1989, Planta (177, 423-434) were used to localize the homogalacturonan (pectate) component of pectins in the cell walls of leaves and soybean root nodules. Leaves of two species of the dicotyledon Dubautia were compared; these species contain much pectin but differ in their tissue water relations with respect to their cell-wall properties. Maturation of the primary cell walls in nodules was studied in the Bradyrhizobium japonicum-Glycine max symbiosis. Probe labelling was based on the divalent-cation-mediated association between pectate in tissue sections and fluorescein-conjugated pectate fragments. Pectate was also labelled by mixed-dimer formation with fluorescent polyguluronate derived from alginate. The specificity of the probe for unesterified polygalacturonate was indicated by increased cell-wall labelling after chemical or enzymatic deesterification of tissue sections, in contrast to elimination of labelling by chemical esterification. Postfixation of tissue sections improved retention of soluble pectate. Pectate differences were found in the leaves among cell types, in degree of esterification, and between plant species. The cell walls of soybean nodules were strongly labelled by the pectate probe in nodules one week and three weeks after infection. Pectate was more highly esterified in the central infected zone than in the surrouding cortex. Within the infected zone, walls of uninfected cells and infected cells were similarly labelled by the pectate probe. The results indicate that the pectate molecular probe provides detailed information on pectate distribution at the cellular level for investigations of cell-wall structure, development and physiology.",
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T1 - Pectate distribution and esterification in Dubautia leaves and soybean nodules, studied with a fluorescent hybridization probe

AU - Vreeland, Valerie

AU - Morse, Suzanne R.

AU - Robichaux, Robert H

AU - Miller, Kathleen L.

AU - Hua, Sui Sheng T

AU - Laetsch, Watson M.

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N2 - Carbohydrate-hybridization probes (Vreeland and Laetsch, 1989, Planta (177, 423-434) were used to localize the homogalacturonan (pectate) component of pectins in the cell walls of leaves and soybean root nodules. Leaves of two species of the dicotyledon Dubautia were compared; these species contain much pectin but differ in their tissue water relations with respect to their cell-wall properties. Maturation of the primary cell walls in nodules was studied in the Bradyrhizobium japonicum-Glycine max symbiosis. Probe labelling was based on the divalent-cation-mediated association between pectate in tissue sections and fluorescein-conjugated pectate fragments. Pectate was also labelled by mixed-dimer formation with fluorescent polyguluronate derived from alginate. The specificity of the probe for unesterified polygalacturonate was indicated by increased cell-wall labelling after chemical or enzymatic deesterification of tissue sections, in contrast to elimination of labelling by chemical esterification. Postfixation of tissue sections improved retention of soluble pectate. Pectate differences were found in the leaves among cell types, in degree of esterification, and between plant species. The cell walls of soybean nodules were strongly labelled by the pectate probe in nodules one week and three weeks after infection. Pectate was more highly esterified in the central infected zone than in the surrouding cortex. Within the infected zone, walls of uninfected cells and infected cells were similarly labelled by the pectate probe. The results indicate that the pectate molecular probe provides detailed information on pectate distribution at the cellular level for investigations of cell-wall structure, development and physiology.

AB - Carbohydrate-hybridization probes (Vreeland and Laetsch, 1989, Planta (177, 423-434) were used to localize the homogalacturonan (pectate) component of pectins in the cell walls of leaves and soybean root nodules. Leaves of two species of the dicotyledon Dubautia were compared; these species contain much pectin but differ in their tissue water relations with respect to their cell-wall properties. Maturation of the primary cell walls in nodules was studied in the Bradyrhizobium japonicum-Glycine max symbiosis. Probe labelling was based on the divalent-cation-mediated association between pectate in tissue sections and fluorescein-conjugated pectate fragments. Pectate was also labelled by mixed-dimer formation with fluorescent polyguluronate derived from alginate. The specificity of the probe for unesterified polygalacturonate was indicated by increased cell-wall labelling after chemical or enzymatic deesterification of tissue sections, in contrast to elimination of labelling by chemical esterification. Postfixation of tissue sections improved retention of soluble pectate. Pectate differences were found in the leaves among cell types, in degree of esterification, and between plant species. The cell walls of soybean nodules were strongly labelled by the pectate probe in nodules one week and three weeks after infection. Pectate was more highly esterified in the central infected zone than in the surrouding cortex. Within the infected zone, walls of uninfected cells and infected cells were similarly labelled by the pectate probe. The results indicate that the pectate molecular probe provides detailed information on pectate distribution at the cellular level for investigations of cell-wall structure, development and physiology.

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KW - Root nodule (cell wall)

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