Studies on the Secretion of Maize Root Cap Slime

Paull, Robert E.; Johnson, C. M.; Jones, Russell L.

doi:10.1104/pp.56.2.300

Cited by 59 publications

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“…The polysaccharide produced by roots of corn contains a high proportion of fucose and galactose, together comprising 69% of the neutral sugars of the polymer (9). Using glucoser4C], Harris and Northcote (3) showed that when the distribution of label appearing in fucose was expressed as a ratio of label in xylose, the ratio was higher in the apical 0.5-mm section of the root.…”

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“…Using glucoser4C], Harris and Northcote (3) showed that when the distribution of label appearing in fucose was expressed as a ratio of label in xylose, the ratio was higher in the apical 0.5-mm section of the root. An autoradiographic study by Kirby and Roberts (7) 20 ,ug/ml of chloramphenicol, and 5 ,ug/ml of streptomycin (9). For Figure 1.…”

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confidence: 99%

“…The alditol acetates were prepared after hydrolysis, as described by Paull et al (9). The neutral sugar alditol acetates were separated on a 183 x 0.318 cm column of 3% ECNSS-M on (100/120 mesh) Gas Chrom Q.…”

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confidence: 99%

“…Then 2 mg of myoinositol were added as a standard and the sample was lyophilized. The sections were hydrolyzed and the alditol acetates were prepared (9).…”

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Studies on the Secretion of Maize Root Cap Slime

Paull

Jones²

1975

Plant Physiol.

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The distribution of fucose-containing polysaccharides in apical 1-cm sections of corn (Zea mays cv. (3) showed that when the distribution of label appearing in fucose was expressed as a ratio of label in xylose, the ratio was higher in the apical 0.5-mm section of the root. An autoradiographic study by Kirby and Roberts (7) Figure 1. Twentyfive root tips and 1 ml of medium were removed from the incubation flask and washed four times with 1 ml of chilled distilled H20. The medium and washings were combined and filtered through a glass fiber disc. The filtrate was made to 80% (v/v) alcohol and allowed to stand at 4 C overnight. The precipitate (secreted material) was then collected on a glass fiber disc and counted. The final soluble filtrate was reduced to a standard volume and is referred to as the supernatant medium. The washed roots were homogenized in 1 ml of chilled glass-distilled H,O in a glass-Teflon homogenizer at 4 C. The brei was either filtered through a glass fiber disc, and the filter was washed with water or centrifuged at 20,000g for 30 min, then resuspended and recentrifuged. The centrifuged pellet or filter disc is referred to as the insoluble cell wall material. The remaining filtrate was combined with the washings, made to 80% (v/v) alcohol, and allowed to stand at 4 C overnight; it was then filtered through a glass fiber disc. The resulting precipitate is designated the homogenate precipitate, and the filtrate, which was reduced in volume, is referred to as unincorporated uptake (Fig. 1).Root tips and secreted materials were also fractionated chemically as outlined by Ray and Baker (10). After the required incubation time in L-fucose-1-[rH], 100 root tips (5 mm) were ground in a glass-Teflon motorized homogenizer at 4 C with 4 ml of incubation medium. The brei was centrifuged at 20,000g for 30 min. The supernatant was retained and the pellet was resuspended with 1 ml of distilled H20 and recentrifuged. The supernatants were combined and the pellet was retained. The supernatant was made 80% (v/v) alcohol and allowed to stand at 4 C for 24 hr. The precipitate was collected on a glass fiber filter disc and is referred to as the cold water-soluble material. The pellet was extracted with 5 307 www.plantphysiol.org on May 10, 2018 -Published by Downloaded from

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confidence: 99%

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confidence: 99%

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confidence: 99%

“…Then 2 mg of myoinositol were added as a standard and the sample was lyophilized. The sections were hydrolyzed and the alditol acetates were prepared (9).…”

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confidence: 99%

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Studies on the Secretion of Maize Root Cap Slime

Paull

Jones²

1975

Plant Physiol.

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“…Their observations were verified and expanded by subsequent workers. The slime from corn was characterized as a high mol wt heteropolysaccharide rich in fucose and galactose (9,17). Secretion and synthesis of slime were studied with high resolution autoradiography (16,19) and by localization of fucose-containing polymers in subcellular fractions from roots incubated in [14CJglucose (2)(3)(4) or [3Hlfucose (20).…”

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Intracellular Localization of GDP-Fucose

James¹,

Jones²

1979

Plant Physiol.

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The intracellular site of synthesis of the fucose-rich polysaccharlde slHme secreted by corn roots was localzed by monitoring the distribution of tions of corn roots. Root tip sectons were chopped in the presence of 0.56 molar sucrose and 100 millmolar Tris (pH 7.0). After a brief centrifugation, the homogenate was appled to a Sepharose 4B column (1.5 x 30 cm). The turbid, particulate portion of the superatant fraction eluted at the void volume. Ninety per cent of the enzyme activity was found in the pooled particulate fractions. The particulate fraction was purified on linear sucrose gradients. Gradient fractions were characterized by buoyant density, 280 nanometer absorbance, electron microscope observation, and distributions of NADH-cytochrome c oxidoreductase and fucosyl transferase activities.Two peaks of fucosyl transferase activity were observed: one in fractions rich in smooth endoplasmic reticulum, and another in fractions rich in dictyosome cisternae and dictyosome vesicles. The data suggest that the synthesis of corn root slime begins in the endoplasmic reticulum and is completed in the dictyosomes prior to secretion; however, the possibility that more than one fucosyl transferase exists in corn roots is discussed.The process of slinie secretion by the peripheral root cap cells of corn was first studied using the electron microscope by Mollenhauer et al. (14). They described large vesicles produced by hypertrophied portions of the dictyosome cisternae that contained electron-dense material that was eventually secreted to the outside of the cell via an exocytotic mechanism. Their observations were verified and expanded by subsequent workers. The slime from corn was characterized as a high mol wt heteropolysaccharide rich in fucose and galactose (9,17). Secretion and synthesis of slime were studied with high resolution autoradiography (16,19) and by localization of fucose-containing polymers in subcellular fractions from roots incubated in [14CJglucose (2)(3)(4) or [3Hlfucose (20).These studies indicate that dictyosomes are involved in the secretion and probably the synthesis of corn root slime.To determine intracellular sites of biosynthesis of the fucoserich slime, organelle fractions were assayed for GDP-fucose:polysaccharide fucosyl transferase activity. This paper describes the organelle fractionation procedure, the results offucosyl transferase localization, and the conclusions that can be drawn from these experiments. MATERIALS AND METHODSPlant Material. Corn seeds (Zea mays cv. SX-17A, a gift of

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Characteristics of Plant Surfaces

Gould

Northcote

1985

Bacterial Adhesion

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Studies on the Secretion of Maize Root Cap Slime

Cited by 59 publications

References 15 publications

Studies on the Secretion of Maize Root Cap Slime

Studies on the Secretion of Maize Root Cap Slime

Intracellular Localization of GDP-Fucose

Characteristics of Plant Surfaces

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