Separation of noncharged carbohydrates from plant material by gas–liquid chromatography of their trimethylsilyl derivatives

Hamlen, R. A.; Lukezic, F. L.; Bloom, J. R.

doi:10.1139/b70-163

Cited by 11 publications

(3 citation statements)

References 6 publications

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“…Evidence from gas chromatographic analysis shows that the exudates contain four sugars including pentoses, the preferred carbon sources for induction of nitrogenase activity (BERGERSEN and GIBSON 1978). Similar studies have revealed that these substances are major constituents of legume root exudates (SHAY and HALE 1973, HAMLEN et al 1970). Evidence from this study is contrary to the conclusion drawn from a plant tissue culture study that the plant cell is capable of supplying the tricarboxylic acid but not pentose sugar (CHILD and KURTZ 1978).…”

Section: Discussionsupporting

confidence: 56%

Root exudates in relation to growth and nitrogenase activity of Rhizobium japonicum

Odunfa

Werner

1981

J of Basic Microbiology

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Root exudates of cowpea and soybean were collected in aerated water cultures. The ability of these exudates t o support nitrogenase activity of R.japonicum was studied on defined media. When a complete nitrogenase-inducing medium was supplemented with concentrated root exudate, there was an increased nitrogenase activity. Whenthecowpea root exudate was substituted for glutamine or sodium succinate in the medium, nitrogenase activity was not detected. Growth was good when t h e root exudate was substituted for sodium succinate but was poor when t h e exudate replaced glutamine. There was nitrogenase activity in R. japonicum when t h e cowpea root exudate was substituted for arabinose in the 20E medium. Gas chromatographic analyses of cowpea root exudates revealed t h a t ribose, arabinose, glucose, and sucrose were the main constituents. The Of the organic substances, a combined nitrogen source, a carbon source and a tricarboxylic cycle intermediate are all that are needed for nitrogenase activity in defined media. Although these substances are common constituents of the root exudates which are the major nutritional source in seedling rhizospheres, there has been no report on the ability of the root exudates to support nitrogen fixation. All that is known so far is that both leguminous and non-leguminous plant tissue cultures supported nitrogenase activity of rhizobia in vitro (CHILD and LARUE 1974, CHILD 1975, WERNER 1976). More recent work shows that diffusible substances from plant tissue cultures in separated systems also enhanced nitrogenase activity (CHILD and KURTZ 1978, STOREY et al. 1979). All these reports establish that the entire requirement for induction of nitrogenase activity in Rhizobium is nutritional. Since the root exudate is the most important nutritional source in the rhizosphere, we decided to investigate the ability of the root exudates to support nitrogenase activity in Rhixobiumjaponicum and to find out if the exudates could replace any of the essential organic nutrients in the nitrogenase-inducing media. Muterials and methodsPlant culture and collection of root exudates: Soybean seeds variety Olima and cowpea seeds variety Ife brown were used. Visually healthy seeds were selected and surface sterilized in 0.1% calcium hypochlorite solution for 5 min. The seeds were subsequently washed in several changes of sterile distilled water and plated on 1% water agar. Cowpea seeds germinated and produced short radicles after 24 hours while soybean took 60 hours to germinate and produce similar lengths of radicles. The seedlings were transferred t o a n exudation apparatus similar to that described by ODUNFA (1979). Each plant culture contained 80 ml of sterile distilled water. The water was aerated 39*

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Section: Discussionsupporting

confidence: 56%

Root exudates in relation to growth and nitrogenase activity of Rhizobium japonicum

Odunfa

Werner

1981

J of Basic Microbiology

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“…The supernatant containing the dissolved sugars was evaporated to dryness. Trimethylsilyl derivatives of the sugars were made by dissolving them in 1.0 ml of pyridine followed by the addition of 0.9 ml of hexamethyldisilazane and 0.1 ml of trifluoroacetic acid (1,6,15,17). Since exudates were recovered from Hoagland's solution, the recovery of a mixture of standard sugars from 3 liters of Hoagland's solution was used to determine the quantitative efficiency of the technique used in this study.…”

Section: Methodsmentioning

confidence: 99%

Effect of Low Levels of Calcium on Exudation of Sugars and Sugar Derivatives from Intact Peanut Roots under Axenic Conditions

Shay

Hale²

1973

Plant Physiol.

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The effects of 10, 20, 35 and 50 mg of Ca2+ per liter on the qualitative and quantitative exudation of sugars from roots of 5-week-old peanut plants, Arachis hypogaea L., grown axenically in nutrient solutions, were measured. Nutrient solutions in which plants had been growing were collected at weekly intervals for 4 weeks, sugars in them were measured by gasliquid chromatography of the trimethylsilyl derivatives. Arabinose, ribose, xylose, fructose, mannose, glucose, galactose, mannitol, galacturonic acid, inositol, sucrose, and five unknowns were found. Qualitative and quantitative differences in exudates were correlated with age of the plants and calcium level. Four times more sugar was exuded at 10 mg than at 50 mg of Ca'+ per liter but no significant differences in growth were observed. Ion efflux measurements suggested that low levels of Ca"' increased root cell membrane permeability.Collection of organic root exudates should be carried out under axenic conditions to prevent losses and alterations by the microbial populations in the rhizosphere. By using the system reported by Hale (4), effects of various factors on qualitative and quantitative exudation of sugars from peanut roots and fruits have been studied (5,10,16). The release of these compounds into the region immediately surrounding roots in soil (rhizosphere) (12) The objectives of the present study were to determine the effect of various low levels of calcium on the root exudation of sugars from axenic peanut plants and to evaluate the possible relationship of exudation to changes in root cell membrane permeability. MATERIALS AND METHODSAseptic peanut seeds, Arachis hypogaea L. cv. Argentine, were obtained by removing the testa and surface sterilizing the remaining part of the seed for 5 min at 50 C in 20% solution of commercial sodium hypochlorite (4). Sterilized seeds were transferred aseptically to the surface of solidified nutrient medium in Petri dishes where they were allowed to germinate at 37 C in a humidified incubator. After 5 days, seedlings were transferred aseptically to sterile, screw-capped, glass jars in which they were transported to each of four sterile isolator chambers (4). The seedlings were transferred from the jars to transplanting tubes containing perlite saturated with Hoagland solution (4). After 4 weeks, 13 sterile (axenic) seedlings per isolator chamber were selected for uniformity and placed in separate 1-liter flasks containing nutrient solution. Random sterility checks of materials inside the chamber, of plant surfaces, and of the inner chamber surfaces were taken at weekly intervals by swabbing representative surfaces. Swabs were placed on potato dextrose agar, nutrient broth, Levine3 EMB agar, Sabouraud dextrose agar, brain heart infusion agar, and thioglycolate medium and incubated at room temperature or at 37 C. Plant sterility was checked before and after each experiment by removing an extra plant from the chamber and macerating samples of leaf, stem, and root tissue, placing them on media, and incubat...

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“…6). In other plants the presence of sorbitol and mannitol may indicate diseased condition (Hamlen, Lukezic & Bloom, 1970) and both may be identified with 5% OV-1 as liquid phase on 60j80 mesh chromosorb. Cereals are of particular interest to the brewing industry and glc has been applied to the determination of sugars in distilled spirits (Martin & Eib, 1968) and in wort (Otter, 1967).…”

Section: Applications To Foodmentioning

confidence: 99%

Gas‐liquid chromatography of food carbohydrates with special reference to trimethylsilyl derivatives A review

Birch

1973

Int J of Food Sci Tech

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Separation of noncharged carbohydrates from plant material by gas–liquid chromatography of their trimethylsilyl derivatives

Abstract: A technique is described for the quantitative separation of very low concentrations of carbohydrates by gas–liquid chromatography using polar and nonpolar silicone liquid phases and TMS (trimethylsilyl) derivatives.

Cited by 11 publications

References 6 publications

Root exudates in relation to growth and nitrogenase activity of Rhizobium japonicum

Root exudates in relation to growth and nitrogenase activity of Rhizobium japonicum

Effect of Low Levels of Calcium on Exudation of Sugars and Sugar Derivatives from Intact Peanut Roots under Axenic Conditions

Gas‐liquid chromatography of food carbohydrates with special reference to trimethylsilyl derivatives A review

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