Food reserves of Scots pine (Pinus sylvestris L.)

Summary The amounts of starch, soluble sugars, triacylglycerols, diacylglycerols and free fatty acids were studied in Scots pine (Pinus sylvestris L.) during an annual cycle in current-year needles and in 1-, 2- and 3-year-old needles collected shortly after bud break. Determination of the compounds was performed using specific enzymatic assays, capillary gas chromatography and thin layer chromatography. Newly emerging needles contained relatively large amounts of starch, but only trace amounts of fat. During autumn and winter, fat content rose, while starch content decreased; amounts of both these reserve materials were very high the next spring shortly before bud break and decreased again during shoot elongation. Concentration of intermediates in triacylglycerol biosynthesis (diacylglycerols and free fatty acids), were low in summer and high in winter. The same pattern was observed for fructose and glucose (the predominant soluble sugars), galactose/arabinose and raffinose/melibiose. In contrast, sucrose concentrations were highest in spring and in autumn. Mature needles of different ages collected in May showed significant differences only in their triacylglycerol and starch content. Concentration changes of reserve materials are discussed in relation to season, mobilization and translocation processes, dormancy, frost resistance and the possibility of carbohydrate-fat interconversions.     

Assessment of sweet potato [Ipomoea batatas (L.) Lam] for bioethanol production in southern Italy

The potential of sweet potato as an alternative crop for bioethanol production has been assessed. We evaluated the amount of soluble sugars, starch and cell wall polysaccharides in tubers of three sweet potato cultivars characterized by different pulp and peel colouration: “Yellow yam” with yellow flesh and brown peel, “White yam” with white flesh and white peel and “Orange yam” with orange flesh and brown peel. The results confirm the high concentration of carbohydrates in sweet potato tubers, especially “Yellow yam”, mainly in the form of starch (67%) and soluble sugars (26%). “Yellow yam”, which is the most widespread cultivar in Salento, appeared the best choice as biomass for bioethanol production. It is characterized by high productivity (20–40 tons/ha year). Results also suggest that “Yellow yam” cultivar has great potential as a bioethanol source in southern Italy with an estimated agroindustrial production yield higher than 2032 l/ha year.     

Regulation of starch and protein accumulation in alfalfa (Medicago sativa L.) somatic embryos

Compared to seeds, somatic embryos accumulated relatively low levels and different types of storage carbohydrates. The regulation of starch accumulation was studied to determine its effects on desiccation tolerance and vigor of dry somatic embryos. Somatic embryos of Medicago sativa are routinely matured through three phases: 7 days of development; 10 days of phase I maturation, a rapid growth phase; and 10 days of phase II maturation, a phase leading to the acquisition of desiccation tolerance. The control of starch deposition was investigated in alfalfa somatic embryos by manipulating the composition of the phase I maturation medium with different levels of sucrose, abscisic acid, glutamine and different types of carbohydrates and amino acids. After phase II maturation, mature somatic embryos were collected for desiccation and subsequent conversion, or for biochemical analyses. Starch deposition occurred primarily during phase I maturation, and variations in the composition of this medium influenced embryo quality, storage protein and starch accumulation. A factorial experiment with two levels of glutamine × three levels of sucrose showed that increasing the sucrose concentration from 30 to 80 g/l increased embryo size and starch content, but had minimal effect on accumulation of storage proteins; glutamine also increased embryo size, but decreased starch content and increased accumulation of the high salt soluble S-2 (medicagin) storage proteins. ABA did not influence any of the parameters tested when included in phase I maturation at concentration up to 10 μM. Replicating sucrose with maltose, glucose, or glucose and fructose did not alter embryo size or starch accumulation (mg/g fresh weight), but replacement with fructose alone reduced embryo size, and replacement with glucose alone reduced germination. Suplementation with the amino acids, asparagine, aspartic acid and glutamine increased seedling vigor, but decreased the starch content of embryos. The data indicate that starch accumulation in somatic embryos is regulated by the relative availability of carbon versus nitrogen nutrients in the maturation medium. The quality of mature somatic embryos, determined by the rate of seedling development (conversion and vigor), correlated with embryo size, storage protein and free amino acid but not with starch. Therefore, further improvements in the quality of somatic embryo may be achieved through manipulation of the maturation medium in order to increase storage protein, but not starch deposition.     

Food reserves of scots pine (Pinus sylvestris L.)

Summary Starch, soluble sugars, triacylglycerols, diacylglycerols and free fatty acids were measured in 30-year-old Scots pine (Pinus sylvestris L.) trees during an annual cycle in the sapwood (youngest ten xylem rings). The radial distribution of carbohydrates and lipids was studied in the trunkwood of 90 -to 150-year-old Scots pine trees collected at the end of the growing season. Determination of the compounds was performed using specific enzymatic assays, capillary gas chromatography and thin layer chromatography. The amounts of glucose, fructose, sucrose, and galactose/arabinose in the sapwood were slightly higher in winter than in summer. Raffinose/stachyose increased up to 5-fold during the cold period. At the beginning of the growing season starch amounts rose, and then decreased in summer and autumn. No concentration changes were observed in the total amounts of diacylglycerols and fatty acids throughout the year. Triacylglycerol levels were slightly higher in February than in summer and autumn. Relative frequencies of individual fatty acids were similar in all lipid fractions. Glucose, fructose, sucrose, starch and triacylglycerols disappeared almost entirely at the transition zone from sapwood to heartwood. In contrast, free fatty acids and galactose/arabinose rose in centripetal direction, and diacylglycerols remained constant across trunk cross-sections. The relative amounts of individual fatty acids changed markedly in the free fatty acid fraction and in the triacylglycerols when crossing the sapwood-heartwood boundary. Concentration changes of reserve materials are discussed in relation to season, mobilization and translocation processes, dormancy, frost resistance, and heartwood formation. The results are compared to those found in needles.     

The influence of zinc and sulphur deficiency on oil-filling in peanut (Arachis hypogaea L.) kernels

In the developing peanut (Arachis hypogaea L.) kernels, the period between 15 and 35 days after podding (DAP) was identified as the active period of oil-filling. The period of active oil-filling was associated with a decrease in the starch, soluble sugars and proteins so as to make available the energy and carbon skeleton for the synthesis of oil. The oil content in the mature kernels decreased by 11, 12 and 25 per cent with Zn, S and Zn+S deficiency, respectively. In addition, proteins and starch content decreased significantly while that of soluble sugars increased slightly. The activity of malate dehydrogenase and glucose-6-phosphate dehydrogenase also decreased due to Zn as well as S deficiency. The deficiency treatments resulted in a decrease in phospholipids, free fatty acids and triacylglycerols in mature kernels. Further the proportion of 16∶0 and 18∶2 decreased while that of 18∶1 increased in developing kernels.     

Mechanisms by which resistant starches and non-starch polysaccharide sources affect the metabolism and disposition of the food carcinogen, 2-amino-3-methylimidazo[4,5-f]quinoline

Although both non-starch polysaccharides (NSP) and resistant starches (RS) are included in current definitions of dietary fibre, our previous work has suggested fundamental differences in the way in which these two classes of material affect the disposition and absorption of a dietary carcinogen. The present studies explore whether different effects on carcinogen metabolism could play a role in the contrasting patterns seen previously. Groups of female Wistar rats were pre-fed for 4 weeks one of five types of defined diet (AIN-76). The control diet contained 35% maize starch and no dietary fibre. The RS-containing diets had all the maize starch substituted with either Hi-maize or potato starch. In the NSP-containing diets, 10% of the maize starch was substituted with dietary fibre in the form of either lignified plant cell walls (wheat straw) or soluble dietary fibre (apple pectin). Pre-fed rats were gavaged with the food carcinogen, [2-14C] 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), and plasma and urinary metabolites characterized using HPLC at various time intervals after administration. After 4 h gavage, plasma from rats on both RS-containing diets contained significantly higher levels of intact IQ and lower levels of the major metabolites, IQ-5-O-glucuronide and IQ-5-sulfate, as compared with plasma from the negative control group at this time. In contrast, plasma from animals on the NSP-containing wheat straw diet (and to a lesser extent the apple pectin diet) showed significantly lower levels of intact IQ, and significantly higher levels of the two major metabolites, as compared with those from the control rats. These different metabolite profiles were also reflected in different urinary excretion profiles. Urine from rats pre-fed RS-containing diets revealed significantly slower metabolite excretion as compared with urine from rats that had been given the NSP-containing diets. Western blotting methodologies also profiled differences between the effects of these two types of dietary fibre in the expression of xenobiotic metabolizing enzymes. We conclude that changes in activity and expression of xenobiotic metabolising enzymes could play a role in the contrasting effects of these two types of dietary fibre on carcinogen uptake and disposition.     

Changes in Total Nitrogen and Free Amino Acids in Stem Cuttings of Mulberry (Morus alba L.)

Changes in total nitrogen and free amino acid contents in stemcuttings of Morus alba have been studied. The fresh and dryweights and total nitrogen amounts of the parent stems of cuttingsdecreased initially after cutting. Their increase follows theformation of main roots in cuttings, suggesting that, like carbohydrates,sugars and starch, stored nitrogenous substances are used forsprouting and rooting of cuttings. Amino acids found in stems,roots and shoots are those common in other higher plants withthe exception of pipecolic acid and 5-hydroxypipecolic acid.Significant changes in the levels of asparagine, proline, arginine,-aminobutyric acid and alanine in roots, bark and wood of parentstems were observed during cutting growth, whereas those ofother amino acids remained comparatively constant; the mostpredominant amino acid in the starting materials was proline.while that in the cuttings during growth was asparagine. Theresults suggest that, among free amino acids, asparagine, prolineand arginine play the major part in storage of nitrogen in mulberry.The importance of glut-amine and asparagine in nitrogen metabolismin mulberry has been discussed.     

ABA associated biochemical changes during somatic embryo development in Camellia sinensis (L.) O. Kuntze

The effect of ABA on reserve accumulation in maturing somatic embryos of tea was compared with and without ABA treatment. Changes in the levels of starch, total soluble sugars (TSS), proteins, and phenols were studied in the somatic embryos at different stages of development (globular, heart, torpedo and germinating embryos) in order to investigate whether ABA could trigger accumulation of storage reserves and thereby overcome the problem of poor germination. After ABA treatment (5.0 mg l(-1)) for 14 days, the starch and protein contents that were negligible in the untreated embryos increased by several fold with a simultaneous increase in TSS. When ABA treatment occurred at the heart stage, the germination of the embryos also improved, relative to untreated controls, after ABA treatment. ABA treatment prior to or after heart stage did not improve somatic embryo germination.     

Characterisation of the cell walls of loquat (Eriobotrya japonica L.) fruit tissues

Loquat fruit (Eriobotrya japonica L. cv. Algor) was dissected to give the following tissue zones: epidermis or epicarp, flesh or mesocarp, integument (a thin layer surrounding the seed cotyledons), seed testa, kernel and hairy receptacle. The alcohol insoluble residues (AIRs) from all these tissues were proved to be free of starch, except loquat kernel which on a fresh weight basis contained about 34% of starch. AIRs were analysed for moisture, ashes, protein, lignin and the component sugars were released by two hydrolytic procedures which helped to distinguish the sugars from non-cellulosic polysaccharides and cellulose. Their major component polysaccharides were inferred to be pectic polysaccharides since all AIRs were very rich in sugars such as uronic acids, arabinose and galactose. Pectic polysaccharides contributed up to 70% of total cell wall polysaccharides in the edible flesh of the loquat fruit. Important differences in the degree of branching, degree of esterification and in the amounts of Ca and Mg associated with pectic polysaccharides were detected among pectic polymers depending on the loquat tissue zone. These compositional and structural differences may be related to the role that these pectic polymers play within the tissues which form the loquat fruit.     

Effects of temperature and photoperiod on postponing bermudagrass (Cynodon dactylon [L.] Pers.) turf dormancy

Growth chamber and field experiments were carried out to determine the effects of extended photoperiod under low and freezing temperatures on bermudagrass turf dormancy at Bajgah, in the southern part of Iran. The experiment in the growth chamber was conducted with four temperature regimes (15, 7.5, 0 and -7.5°C) and three light durations (8, 12 and 16h) in a completely randomized design with four replications. The field study was conducted in two consecutive years (2008-2009) with three light durations (8, 12 and 16h) in months with natural short day length and arranged in a randomized complete blocks design with three replications. Results in both experiments showed that decreasing temperature and photoperiod decreased verdure fresh and dry weight, shoot height, tiller density, leaf area and chlorophyll and relative water contents (RWC). However, rooting depth and fresh weight of roots increased in the growth chamber. Decreasing the temperature and light duration increased electrolyte leakage and proline content. Reducing sugars increased with decreasing temperature and declined with lowering light duration in both shoots and roots. Starch content of both shoots and roots showed an adverse trend compared to reducing sugars; starch content increased in both shoots and roots in all treatments by shortening the photoperiod. Practically, the problem of bermudagrass turf's dormancy could be solved via increasing the photoperiod in months with short day lengths. This treatment would be efficient and useful for turfgrass managers to apply in landscapes and stadiums.     

Comparative proteomic analysis of cold-induced sweetening in potato (Solanum tuberosum L.) tuber

Cold-induced sweetening is one of the major factors limiting the quality of fried potato products. To understand the mechanisms of protein regulation for cold-induced sweetening in potato tubers, a comparative proteomic approach was used to analyse the differentially expressed proteins both during control (25 °C, 30 days) and cold treatment (4 °C, 30 days) using two-dimensional gel electrophoresis. Quantitative image analyses indicated that there were 25 protein spots with their intensities significantly altered more than twofold. Of these proteins, 9 were up-regulated, 13 were down-regulated, 2 were absent, and 1 was induced in the cold-stored tubers. The MALDI-TOF/TOF MS analyses led to the identification of differentially expressed proteins that are involved in several processes and might work cooperatively to maintain metabolic homeostasis in tubers during low-temperature storage. The preponderance of metabolic proteins reflects the inhibition of starch re-synthesis and the accumulation of sugars in carbon fluxes, linking starch–sugar conversion. The respiration-related proteins suggest the transfer of respiratory activity from aerobic respiration to anaerobic respiration in the cold-stored tubers. The proteins associated with defence appear to protect the tuber cells from low-temperature stress. Some heat shock proteins that act as chaperones also displayed a differential expression pattern, suggesting a potentially important role in cold-stored tubers, although their exact contribution remains to be investigated. The proposed hypothetical model might explain the interaction of these differentially expressed proteins that are associated with cold-induced sweetening in tubers.     

Evidence of the crucial role of sucrose synthase for sink strength using transgenic potato plants (Solanum tuberosum L.)

Sink strength of growing potato tubers is believed to be limited by sucrose metabolism and/or starch synthesis. Sucrose synthase (Susy) is most likely responsible for the entire sucrose cleavage in sink tubers, rather than invertases. To investigate the unique role of sucrose synthase with respect to sucrose metabolism and sink strength in growing potato tubers, transgenic potato plants were created expressing Susy antisense RNA corresponding to the T-type sucrose synthase isoform. Although the constitutive 35S CaMV promotor was used to drive the expression of the antisense RNA the inhibition of Susy activity was tuber-specific, indicating that independent Susy isoforms are responsible for Susy activity in different potato organs. The inhibition of Susy leads to no change in sucrose content, a strong accumulation of reducing sugars and an inhibition of starch accumulation in developing potato tubers. The increase in hexoses is paralleled by a 40-fold increase in invertase activities but no considerable changes in hexokinase activities. The reduction in starch accumulation is not due to an inhibition of the major starch biosynthetic enzymes. The changes in carbohydrate accumulation are accompanied by a decrease in total tuber dry weight and a reduction of soluble tuber proteins. The reduced protein accumulation is mainly due to a decrease in the major storage proteins patatin, the 22 kDa proteins and the proteinase inhibitors. The lowered accumulation of storage proteins is not a consequence of the availability of the free amino acid pool in potato tubers. Altogether these data are in agreement with the assumption that sucrose synthase is the major determinant of potato tuber sink strength. Contradictory to the hypothesis that the sink strength of growing potato tubers is inversely correlated with the tuber number per plant, no increase in tuber number per plant was found in Susy antisense plants.     

Changes in protein and carbohydrate content during development of seeds and siliquas of rapeseed (Brassica napus L.)

Abstract

A study was made on the changes observed in the protein, starch and soluble sugar content during development of siliquas and seeds of rapeseed grown in central Italy.

Concentration of starch and soluble sugars in the seed increases to 75 per cent dry matter during the first few weeks of pod development and then drops to minimum values. The protein increases steadily until maturity, when a level of 0.85 mg per seed is reached, equivalent to 18 per cent dry matter. The protein and starch in the hull? decrease continuously during development, while in the initial stages the soluble sugars are accumulated until they account for 33 per cent dry matter, after which they decline towards maturity.     

Sugar metabolism and developmental stages of rubber tree (Hevea brasiliensis L.) seeds

Changes in the concentration of sugars and sucrose metabolism enzymes can characterize the developmental stages of a seed. In recalcitrant species such as Hevea brasiliensis L., little is known about these changes. We aimed to evaluate the three main stages of development of rubber tree seeds – histodifferentiation, cell elongation and accumulation of reserves. The activities of acid and neutral invertases (E.C. 3.2.1.26) and sucrose synthase (EC 2.4.1.13), and the concentrations of reducing sugars (RS), total soluble sugars (TSS) and sucrose (Suc) were determined concomitantly with the histochemical and anatomical evaluation of seed structure. Histodifferentiation in rubber tree seeds occurs up to 75 days after anthesis (DAA). The concentration of RS is high and of Suc is low during seed histodifferentiation, which occurs along with a visible increase in the number of cell divisions. After that period, there is an increase in the concentration of Suc (mg g^?1) and in the number and size of starch granules, and a decrease in the concentration of RS (mg g^?1). At that point, cell elongation occurs. At 135 DAA, there is an inversion in the concentration of these two sugars and an increase in reserve accumulation. Thus, in seeds of the evaluated clone, the period up to 75 DAA is characterized as the histodifferentiation stage, while from that time up to 120 DAA the cell elongation stage takes place. The final stage of seed maturation and reserve accumulation begins at 135 DAA, and the seed, including the embryo, is completely formed at 175 DAA.     

Changes in Carbohydrate Levels in Red Kidney Bean (Phaseolus vulgaris L.) Exposed to Sulphur Dioxide

Red kidney bean (Phaseolus vulgaris L.) was exposed to variousconcentrations of SO² for 24 h (16 h light/8 h dark photoperiod)with continuous monitoring of photosynthetic and respiratoryactivity. Plants were harvested at the end of the dark periodinto samples of mature and immature leaf tissue, stems, androots for determination of sugar and starch levels. In all tissue samples the levels of total sugars were increasedby exposure to the lower concentrations of SO², but decreasedby the higher concentrations. Starch levels in leaves followeda similar trend. Increases in sugar and starch levels precededsymptoms of visible injury. Decreasing rates of photosynthesiswere correlated with increasing rates of respiration, the occurrenceof visible injury, and the depletion of sugar and starch levels.     

Long-term Metabolism of [14C]Sugars in Stored Potatoes

[¹⁴C]Sucrose, [¹⁴C]glucose and [¹⁴C]fructose were introducedinto potato tubers held at 10 °C and the redistributionof label chased over a 65 d period in storage. Respiratory losseswere identical in all treatments, as was the partitioning of¹⁴C between soluble and insoluble forms. Sucrose was the predominantlabelled sugar in the tubers after 20 h, regardless of the original[¹⁴C]sugar introduced, and was loaded and distributed throughoutthe tubers by the internal phloem system. After 20 h the proportionsof labelled sugars bore no relationship to those of the unlabelledendogenous sugars. However, with time the percentage of ¹⁴Cin sucrose fell while that in glucose increased and by 65 dthe proportions of the labelled sugars more closely resembledthe endogenous pools. Fructose represented a consistently lowproportion of both the labelled and unlabelled sugars. By 21d a considerable proportion of the soluble ¹⁴C had been convertedto starch (approx. 25% of the total tuber 14C), this value remainingrelatively constant for the remainder of the storage period.Sprouts which formed on the tubers contained up to 6% of thetotal tuber ¹⁴C but less than 0.2% of the tuber dry matter.It is suggested that the bulk of the translocated [¹⁴C]sucroseentered the symplast and exchanged slowly with the bulk of thesugars in the storage cell vacuoles. [¹⁴C]sugars, phloem loading, starch, potato tuber, Solunum tuberosum, cold storage     

Changes of Carbohydrates in Pepper (Capsicum annuum L.) Flowers in Relation to Their Abscission Under Different Shading Regimes

Abscission of pepper flowers is enhanced under conditions oflow light and high temperature. Our study shows that pepperflowers accumulate assimilates, particularly in the ovary, duringthe day time, and accumulate starch, which is then metabolizedin the subsequent dark period. With the exception of the petals,the ovary contains the highest total amounts of sugars and starch,compared with other flower parts and contains the highest totalactivity, as well as activity calculated on fresh mass basis,of sucrose synthase, in accordance with the role of this enzymein starch biosynthesis. Low light intensity or leaf removaldecreased sugar accumulation in the flower and subsequentlycaused flower abscission. The threshold of light intensity fordaily sugar accumulation in the sink leaves was much lower thanin flowers, resulting in higher daytime accumulation of sugarsin the sink leaves than in the adjacent flower buds under anylight intensity, suggesting a competition for assimilates betweenthese organs. Flowers of bell pepper cv. ‘Maor’and ‘899’ (sensitive to abscission) accumulatedless soluble sugars and starch under shade than the flowersof bell pepper cv. ‘Mazurka’ and of paprika cv.‘Lehava’ (less sensitive). The results suggest thatthe flower capacity to accumulate sugars and starch during theday is an important factor in determining flower retention andfruit set. Pepper; Capsicum annuum L.; abscission; shading; pepper flowers; ovary; leaves; sugars; starch; acid invertase; sucrose synthase