Extraction of Total Available Carbohydrates from Grass and Legume Tissue

Smith, Dale; Paulsen, Gary M.; Raguse, C. A.

doi:10.1104/pp.39.6.960

Cited by 227 publications

(150 citation statements)

References 13 publications

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“…Lipids were extracted with chloroformmethanol (2: 1, v/ v) from frozen materials and estimated gravimetrically. Insoluble nonstructural carbohydrate was estimated by the anthrone method in lipid-free residue hydrolyzed with 0.2 N H2SO0 (23).…”

Section: Methodsmentioning

confidence: 99%

Adenylate Energy Pool and Energy Charge in Maturing Rape Seeds

Ching¹,

Crane²,

Stamp³

1974

Plant Physiol.

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A study of energy state and chemical composition of pod walls and seeds of maturing rape (Brassica napus L.) was conducted on two varieties, Victor anid Gorezanski. Total adenosine phosphates, ATP, and adenylate energy charge increased with increasing cell number and cellular synthesis during the early stages, remained high at maximum dry weight accumulation and maximum substrate influx time, and decreased with ripening. A temporal control of energy supply and ATP concentration is evident in developing tissues with determined functions; whereas the association of a high energy charge and active cellular biosynthesis occurs only in tissues with a stabilized cell number.Maturing seeds require a tremendous supply of ATP for the biosynthesis of various seed components, for the biochemical work, such as the loading and transport of phloem sap and minerals and the intracellular cytoplasmic streaming, and particularly for the accumulation of RNAs with the increasing number of cells and cellular activities. It is of interest to measure this energy supply during seed development and maturation and to discern whether or not a regulatory role of the adenylate energy charge is imposed on these processes. MATERIALS AND METHODSMaterials. Two varieties, Victor and Gorczanski, of rape (Brassica napus L.) were planted at a rate of 1.2 kg/ha in a 10-cm row spacing at Corvallis, Oregon, on September 28, 1972. The soil was a silty clay loam with a pH of 5.0 to 5.4. A preplant fertilizer application of 400 kg of N/ha and an additional 100 kg of N/ha on March 6, 1973 were applied. Weed control was effected by preplant incorporation of 4 kg/ha of Eptam (S-ethyl dipropylthiocarbamate) and 3.4 kg/ha of Treflan (a, a, a-trifluoro-2, 6-dinitro-N'N-dipropyl-p-toluidine).No irrigation was applied because the natural rainfall was enough to provide the water needs of the plants. On Sample collection. Two representative racemes were selected for each sampling date for each variety. The pods on one side of the raceme were cut off and immediately dropped into liquid nitrogen for chemical analyses in the laboratory. Because of the rapid change in the contents of adenosine phosphates in tissues, liquid N2 was used to maintain their in situi content during transport. The other half of the pods were kept in polyethylene bags in crushed ice for the determination of weight, size, and water content. The time of sample collection was consistently maintained around 9:00 AM to minimize diurnal variation.Extraction of Adenosine Phosphates (AP). Pods in liquid N2 were poured into a supercooled aluminum pan, packed in Dry Ice. The pods were classified into size classes of 1-cm increments and the number of pods in each class was recorded. Pods smaller than 5 cm in length were extracted whole because the seeds were inseparable from the pod wall. Larger pods were dissected to pod walls and seeds under a frozen condition, then extracted separately. At least two replications were analyzed for each size class and a total of 8 to 20 analyses were conducte...

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Section: Methodsmentioning

confidence: 99%

Adenylate Energy Pool and Energy Charge in Maturing Rape Seeds

Ching¹,

Crane²,

Stamp³

1974

Plant Physiol.

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“…Protein was extracted from replicate 10mg samples with 0.1moll -1 NaOH and determined using the Bio-Rad Protein Microassay for Microtiter Plates based on the Bradford assay (Bio-Rad Laboratories, Hercules, CA, USA). Total non-structural carbohydrate was determined colorimetrically from 15mg replicate samples following extraction with 0.1moll -1 H 2 SO 4 (Smith et al, 1964) using the phenol-sulphuric assay (DuBois et al, 1956). Carcass lipid was determined gravimetrically following three 24h extractions with 100% chloroform.…”

Section: Locusts and Dietsmentioning

confidence: 99%

Insect herbivores can choose microclimates to achieve nutritional homeostasis

Clissold

Coggan

Simpson

2013

Journal of Experimental Biology

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SUMMARYThe interaction between temperature and diet quality can affect the life history of ectotherms. The rate and ratio at which protein and carbohydrate are obtained from food are important aspects of diet quality, and insects have a well-developed capacity to adjust their feeding behaviour and post-ingestive physiology to regulate intake and allocation of these nutrients. If the supply of protein and carbohydrate varies with temperature (e.g. via effects on intake, digestion or metabolism), then herbivorous insects can use thermoregulatory behaviour to help achieve nutritional homeostasis. When fed the host grass Triticum aestivum, Locusta migratoria nymphs absorbed and allocated protein and carbohydrate to growth with the same efficiency at 38°C as at 32°C; however, at the higher temperature, they ingested more food. In contrast, when feeding on Themeda triandra, the nymphs absorbed carbohydrate with higher efficiency at 32°C, and protein at 38°C. Using synthetic diets, we induced either a protein or a carbohydrate deficiency in experimental insects and showed that locusts placed in a thermal gradient following a meal of T. triandra selected 32°C when deprived of carbohydrate, and 38°C when deprived of protein. This capacity to use thermoregulatory behaviour to redress an imposed nutritional imbalance improved with experience of feeding on T. triandra. As predicted, locusts fed T. aestivum always chose higher temperatures, irrespective of nutritional state. Our results have consequences for understanding host-plant choice by herbivores and interpreting the effects of changed environmental temperatures and microclimate on animal-plant interactions. Supplementary material available online at

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“…Protein was extracted from replicate 10mg samples of plant and faecal material with 0.1moll -1 NaOH and determined using the Bio-Rad micro assay (Bio-Rad, Hercules, CA, USA) based on the Bradford assay. Total non-structural carbohydrate was determined colourimetrically from 10mg plant and faecal samples following extraction with 0.1moll -1 H 2 SO 4 (Smith et al, 1964), using the phenol-sulphuric assay (DuBois et al, 1956). Neutral detergent fibre was determined gravimetrically from replicate 50mg plant samples using the Van Soest method, omitting sodium sulphite (Van Soest et al, 1991;Clissold et al, 2004).…”

Section: Determination Of Diet Composition and Nutrient Absorption Frmentioning

confidence: 99%

Protein-induced weight increase of the gastrointestinal tract of locusts improves net nutrient uptake via larger meals rather than more efficient nutrient absorption

Clissold

Brown

Simpson

2012

Journal of Experimental Biology

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SUMMARYIncreasing the tissue biomass and/or volume of the gastrointestinal tract (GIT) is commonly seen when animals feed on poorquality diets. This increase can simply permit larger meal sizes, but may also rebalance nutritionally imbalanced ingesta by allowing selective absorption of limiting nutrients. In an insect herbivore, the migratory locust, a synthetic diet with a high ratio of protein to carbohydrate was found to induce mass enhancement of the GIT. When normalised for sex and overall body size, increases to the mass of the foregut and midgut caeca resulted in higher absorption (20-30%) of both protein and carbohydrate when subsequently feeding on three chemically and structurally different grasses. Greater net absorption of macronutrients occurred because these locusts ate larger meals that transited at the same time and with the same digestive efficiency as locusts in which the GIT was not enlarged. Thus, plasticity of the GIT did not improve nutritional homeostasis, but increased the rate of nutrient uptake. Supplementary material available online at

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Extraction of Total Available Carbohydrates from Grass and Legume Tissue

Cited by 227 publications

References 13 publications

Adenylate Energy Pool and Energy Charge in Maturing Rape Seeds

Adenylate Energy Pool and Energy Charge in Maturing Rape Seeds

Insect herbivores can choose microclimates to achieve nutritional homeostasis

Protein-induced weight increase of the gastrointestinal tract of locusts improves net nutrient uptake via larger meals rather than more efficient nutrient absorption

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