Staphylococci in Competition: III. Influence of pH and Salt on Staphylococcal Growth in Mixed Populations

Previous results showed definite repressive effects on the growth of staphylococci in mixed cultures due to the competitive growth of psychrophilic saprophytes. This study was continued, and the influence of other environmental factors, pH and salt, on the competition between staphylococci and saprophytes was investigated. Initial pH values varied from 5 to 9. At the extremes of the pH range, staphylococci failed to grow, while the saprophytes grew under all of the conditions tested. At pH 5, the growth curves for the saprophytes were markedly altered from those obtained at neutral pH. The lag phases were greatly lengthened at and below 20 C, but normal numbers of saprophytes were reached in the stationary phase. At pH 6 and 8, staphylococcal growth showed the same inhibition observed at pH 7, at and below 20 C; normal multiplication was observed above this temperature, but with accelerated death phases. Thus, pH did not primarily effect staphylococcal growth through its influence on saprophyte growth and competition, but rather directly affected the growth of Staphylococcus cultures. Salt concentrations from 3.5 to 9.5% were investigated for influence on staphylococcal growth in mixed populations. Above 3.5% salt, staphylococcal inhibition at and above 20 C was not as marked as in the controls, although normal numbers were never reached. The saprophytes were increasingly inhibited, and their lag phases materially lengthened as salt concentration was increased. Salt acted directly on the Staphylococcus population and also, by repressing saprophyte growth, decreased competition, which allowed the staphylococci to grow.     

Staphylococci in Competition: V. Effect of Eggs, Eggs Plus Carbohydrates, and Lipids on Staphylococcal Growth

As is generally known, custards have been frequently involved in staphylococcal food poisonings and are regarded by some as an ideal culture medium. Previous studies showed that high carbohydrate concentrations repressed the growth of competing saprophytic species, allowing the growth of staphylococci in a mixture rather than stimulating the growth of the staphylococci. In an extension of those studies, the influence of the egg constituent of custard was investigated to determine its role in affecting the growth of pathogenic staphylococci in a competing mixture of psychrotrophic saprophytes. The normal competitive effect of the saprophytes on the growth of staphylococci was very slightly affected by the addition of 25% whole egg to the growth media. Approximately 9% egg yolk alone added to the medium resulted in a slight increase in the length of the lag period of the psychrotrophs and a slight increase in the number of staphylococci which grew. Addition of 25% whole egg plus 14.5% sucrose resulted in repression of saprophyte growth similar to that seen in high sucrose concentrations. Staphylococcal growth was more extensive in the presence of both whole egg and sucrose than in the presence of either ingredient alone. Incorporation of 4% corn oil in media was effective in repressing growth of the saprophytes at 37 C only. This allowed the staphylococci to dominate the population. At lower temperatures, staphylococci were unable to compete effectively. Buffering media of high carbohydrate content resulted in lengthened lag periods for the psychrotrophs and the appearance of very large staphylococcal populations.     

Staphylococci in Competition: II. Effect of Total Numbers and Proportion of Staphylococci in Mixed Cultures on Growth in Artificial Culture Medium

In studies carried on in bacteriological media with selected cultures, definite repressive effects were noted on the growth of the Staphylococcus population by a mixture of saprophytic, psychrophilic bacterial species. This repressive effect became more pronounced as the relative proportion of the bacterial population which was staphylococcal became smaller. A varied saprophytic bacterial flora of some numbers apparently would offer definite protection to foods through repression of staphylococcal growth and by rendering the food inedible before the rise of appreciable numbers of staphylococci. It would appear that at the optimal temperature for staphylococcal growth, staphylococci could multiply rapidly in the mixed population due to the comparative shortness of the generation time of this species and because of the lengthened lag phase of the saprophytic bacterial species at this elevated temperature, especially when only cultures having psychrophilic characteristics were present. This temperature is substantially above that encountered in practical experience. With the passage of time, the staphylococcal population was completely overgrown by the saprohytes present. This effect might be eliminated in the presence of psychrophilic and mesophilic, saprophytic species. The repressive effect of competition by saprophytic, psychrophilic organisms is extremely effective up to room temperature on the staphylococcal population. Even when significant staphylococcal populations were achieved in the artificial media, such tremendous numbers of saprophytes were obtained either earlier or at the same time so that a frozen food containing this population would be organoleptically unacceptable due to the degradative action of enzymes from the saprophytic psychrophile population.     

Influence of Food Microorganisms on Staphylococcal Growth and Enterotoxin Production in Meat

Forty-four microorganisms were studied for their influence on staphylococcal growth and enterotoxin production. Inhibition was found to be more common than stimulation. Two types of inhibition were observed: inhibition of staphylococcal growth, and inhibition of enterotoxin formation with no apparent effect on growth. By use of a plate test, 12 of the 44 food microorganisms were found to inhibit staphylococcal growth at 35 C. Of the 12, 3 also inhibited growth at 25 C. No significant differences in inhibition were observed with the 15 strains of enterotoxigenic staphylococci. In meat slurries, inhibition of staphylococcal growth was found to be greater at 25 C than at 35 C. Results on inhibition obtained from the plate test could not be correlated with the effect of the organisms in slurries. Environmental conditions were found to affect markedly the influence of food microorganisms on staphylococci. Of the 44 food microorganisms studied, only Bacillus cereus was observed to stimulate significantly staphylococcal growth and enterotoxin formation. Stimulation was more pronounced with Staphylococcus aureus 196E than with other strains of enterotoxigenic staphylococci. Bacillus megaterium and Brevibacterium linens were inhibited by staphylococci. These organisms were completely inhibited when inoculated in mixed cultures with staphylococci. In pure cultures, good staphylococcal growth was found to be accompanied by enterotoxin production; however, in the presence of food microorganisms, good staphylococcal growth occurred without the formation of detectable levels of enterotoxin A.     

Excess nickel modulates activities of carbohydrate metabolizing enzymes and induces accumulation of sugars by upregulating acid invertase and sucrose synthase in rice seedlings

The effects of increasing concentrations of nickel sulfate, NiSO₄ (200 and 400 μM) in the growth medium on the content of starch and sugars and activity levels of enzymes involved in starch and sugar metabolism were examined in seedlings of the two Indica rice cvs. Malviya-36 and Pant-12. During a 5–20 day growth period of seedlings in sand cultures, with Ni treatment, no definite pattern of alteration in starch level could be observed in the seedlings. In both roots and shoots of the seedlings Ni treatment led to a significant decrease in activities of starch degrading enzymes α-amylase, β-amylase, whereas starch phosphorylase activity increased. The contents of reducing, non-reducing, and total sugars increased in Ni-treated rice seedlings with a concomitant increase in the activities of sucrose degrading enzymes acid invertase and sucrose synthase. However, the activity of sucrose synthesizing enzyme sucrose phosphate synthase declined. These results suggest that Ni toxicity in rice seedlings causes marked perturbation in metabolism of carbohydrates leading to increased accumulation of soluble sugars. Such perturbation could serve as a limiting factor for growth of rice seedlings in Ni polluted environments and accumulating soluble sugars could serve as compatible solutes in the cells under Ni toxicity conditions.     

The Carbohydrate Nutrition of Tomato Roots: VI. The Inhibition of Excised Root Growth by Galactose and Mannose and its Reversal by Dextrose and Xylose

The addition of 0.01–0.015 per cent. galactose or 0.005–0.01per cent, mannose reduces by 50 per cent, the linear growthof excised tomato roots cultured in a I per cent, sucrose medium.An addition of 0.03–0.04 per cent, of either sugar causesnot less than a 90 per cent, inhibition of growth. The survivalof meriste-matic activity is higher in presence of fully inhibitoryconcentrations of mannose than of galactose. Roots inhibitedby galactose are distinguishable from those inhibited by mannose. The inhibitory effect of concentrations of galactose up to 0.15per cent, and of mannose up to 0.4 per cent, can be fully antagonizedby the simultaneous addition to the culture medium of dextrose.The minimum ratio of dextrose: inhibitory sugar for maximumantagonism of the growth inhibition is with galactose 5: 1 andwith mannose 3.5: 1. Growth of roots in a dextrose-containingmedium does not protect them from subsequent inhibition by eithergalactose or mannose. d-xylose has significant activity as an antagonist of mannoseinhibition and even more so of galactose inhibition. However,the restoration of growth achievable from the addition of xyloseis not comparable with that resulting from the addition of dextrose.The inhibition of growth by xylose is not alleviated by thesimultaneous addition of dextrose. Maltose has low activityas an antagonist of galactose and mannose inhibition. All othersugars tested and the sugar alcohols corresponding to galactoseand mannose were quite inactive as antagonists of the growthinhibition by these two sugars. Mixtures of partially inhibitory concentrations of galactoseand mannose were less inhibitory than their more inhibitorycomponent. The concentration of dextrose required to reversethe inhibitory activity of such mixtures was not greater thanthe minimum concentration required to antagonize the actionof the more inhibitory component. The antagonism of galactose inhibition by dextrose is not dueto dextrose impeding galactose uptake.     

Sugar uptake and starch biosynthesis by slices of developing maize endosperm

¹⁴C-Sugar uptake and incorporation into starch by slices of developing maize (Zea mays L.) endosperm were examined and compared with sugar uptake by maize endosperm-derived suspension cultures. Rates of sucrose, fructose, and d- and l-glucose uptake by slices were similar, whereas uptake rates for these sugars differed greatly in suspension cultures. Concentration dependence of sucrose, fructose, and d-glucose uptake was biphasic (consisting of linear plus saturable components) with suspension cultures but linear with slices. These and other differences suggest that endosperm slices are freely permeable to sugars. After diffusion into the slices, sugars were metabolized and incorporated into starch. Starch synthesis, but not sugar accumulation, was greatly reduced by 2.5 millimolar p-chloromercuribenzenesulfonic acid and 0.1 millimolar carbonyl cyanide m-chlorophenylhydrazone. Starch synthesis was dependent on kernel age and incubation temperature, but not on external pH (5 through 8). Competing sugars generally did not affect the distribution of ¹⁴C among the soluble sugars extracted from endosperm slices incubated in ¹⁴C-sugars. Competing hexoses reduced the incorporation of ¹⁴C into starch, but competing sucrose did not, suggesting that sucrose is not a necessary intermediate in starch biosynthesis. The bidirectional permeability of endosperm slices to sugars makes the characterization of sugar transport into endosperm slices impossible, however the model system is useful for experiments dealing with starch biosynthesis which occurs in the metabolically active tissue.     

The Carbohydrate Nutrition of Tomato Roots: VII. Sugars, Sugar Phosphates, and Sugar Alcohols as Respiratory Substrates for Excised Roots

The respiratory responses of substrate-depleted excised roottips to a range of sugars, sugar phosphates, and sugar alcoholshave been determined by measuring oxygen uptake by the directmethod of Warburg. Sucrose, dextrose, and laevulose are the only sugars which promotea high level of oxygen uptake. The effects of azide and DNP on the oxygen uptake promoted bysucrose and by dextrose are described. Mannose is a strong inhibitor of respiration. This inhibitionis reversed by the simultaneous addition of those sugars whichalso reverse the growth inhibition caused by mannose. Mannoseinhibits the respiration of sucrose and of glycolytic intermediates.Galactose is slowly respired and does not, even at high concentration,inhibit the respiration of sucrose. The results are discussed in relation to the growth effectsof the sugars tested.     

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     

The Growth of Acer pseudoplatanus Cells in a Synthetic Liquid Medium: Response to the Carbohydrate, Nitrogenous and Growth Hormone Constituents

A synthetic culture medium which supports a high level of growth of a scrially propagated cell suspension culture of Acer pseudoplatanus is described. The sucrose of this medium can be effectively replaced by glucose or fructose or a mixture of glucose and fructose or galactose or maltose or soluble starch. When the carbohydrate is glucose or fructose no other sugars appear in the culture medium in significant amounts. Glucose is absorbed in greater quantity than fructose from an equimolar mixture of these sugars. When sucrose is supplied both glucose and fructose appear in the medium. Glucose appears in maltose medium, and maltose and glucose in soluble starch medium. Under the standard conditions of culture, media containing 2 % sucrose or 2 % glucose become depleted of sugar before the 25th day of incubation. Enhanced yield of the cultures can be obtained by raising the initial sucrose concentration to 6 %. – A supply of nitrate is essential for maximum yield and healthy growth. Growth, in the presence of nitrate, is significantly enhanced by a supply of urea. Addition of casein hydrolysate or of a mixture of amino acids enhances growth in the presence of nitrate and urea and particularly when nitrate is omitted. – When kinetin is omitted or incorporated at the standard level (0.25 mg/I), 2,4-dichlorophenoxyacetic acid (2,4-D) at 1.0 mg/l is essential for continuation of growth at a high level. It cannot be replaced by indol-3yl-acetic acid (IAA). 1-naphthaleneacetic acid (NAA) at 10 mg/l permits of a low level of growth with abnormal aggregation. When the level of kinetin is raised to 10 mg/l a high level of growth occurs in the absence of added auxin but the cultures become brown and tend to show increasing aggregation on subculture.     

Endogenous Sugar Levels and Their Effects on Root Formation and Petiole Yellowing of Detached Mustard Cotyledons

Detached cotyledons of Sinapis alba rooted readily in water in petri dishes in the light. The addition of (6 × 10^?2M) galactose, mannose or 2-deoxy-D-glucose to the culture medium proved toxic to cotyledon growth. Of the other sugars tested that were not toxic, sucrose was the most inhibitory to root formation and increased petiolar yellowing to the greatest extent. Glucose was more inhibitory than fructose which in turn increased petiolar yellowing more than methyl-D-glucose. Sucrose, glucose, or fructose at 6 × 10^?2M in the culture medium gave rise to very substantial increases in the cotyledon petiole of reducing sugar and starch with smaller increases in sucrose. Methyl-D-glucose had much less effect on internal sugar levels. It was found that the higher the internal level of glucose the more rapid the rate and final extent of petiolar yellowing. In general, the degree of petiolar yellowing was inversely related to the ability of the cotyledon to root. Methyl-D-glucose differed from the other sugars in that it delayed and reduced root formation but had very little effect on petiolar yellowing.     

The Carbohydrate Nutrition of Tomato Roots: V. The Promotion and Inhibition of Excised Root Growth by Various Sugars and Sugar Alcohols

Iron is only consistently present in an available form in White'sroot culture medium if the inorganic salts are autoclaved withthe sugar. The substitution of ferric ethylenediamine-tetra-acetatefor the inorganic ferric salt of White's medium ensures ironavailability when the carbon source of the medium is renderedsterile by ether treatment and subsequently added to the remainderof the constituents which have been sterilized by autoclaving. The biological activity of sugars, and particularly of dextroseand laevulose, is altered by autoclaving them in presence ofthe inorganic salt solution of White's medium. The only sugar which supports a considerable growth of excisedtomato roots is sucrose. The activity of this sugar is not affectedby alcohol-precipitation nor is it dependent upon the simultaneouspresence of traces of its constituent mono-saccharides. Dextrose or laevulose or a mixture of the two sugars supporta low but sustained level of excised-root growth. All othersugars and sugar alcohols tested were inactive as carbon sources. The addition of sucrose at low concentration (0–2 percent.) to a medium containing dextrose as the main carbon compounddoes not make possible a level of growth comparable with thatobtained with an adequate sucrose supply. It has not been possibleto enhance the growth-rate of excised roots supplied with dextroseby previous presentation of this sugar under conditions permittingactive growth. Using media containing 'etherized' sucrose anddextrose, no evidence was obtained of any competitive inhibitionof sucrose utilization by dextrose. Certain sugars when added to a medium, containing the optimumconcentration of sucrose₁, markedly inhibited excised root growth.Thus l-sorbose, l- and d-arabinose, and d-xylose caused notless than 80 per cent, inhibition at a concentration of 0-5per cent. d-mannose and d-galactose completely inhibited growthat o-1 per cent. The oligosaccharides, dextrose, laevulose,and the sugar alcohols tested had, by contrast, very low inhibitoryactivity.     

Studies on growth and metabolism of Oenococcus oeni on sugars and sugar mixtures

AIMS: To study the effect of sugars and sugar mixtures on the growth kinetics of Oenococcus oeni NCIMB 11648 in batch culture with the aim of producing a high cell productivity system for starter cultures. METHODS AND RESULTS: The growth of O. oeni was investigated on single sugars (glucose, fructose or sucrose) and their mixtures (glucose-fructose, glucose-sucrose or fructose-sucrose). Better growth was obtained on sugar mixtures compared with growth on a single sugar. The production system of O. oeni biomass was investigated in batch culture with or without pH control with respect to kinetics, specific growth rate and biomass yield. The effect of pH and substrate concentration on fermentation balances and ATP yield were determined. The optimal growth of O. oeni was achieved on the glucose-fructose mixture (9 g l(-1), 1 : 1) at pH 4.5 and 25 degrees C with pH control, with highest cell volumetric productivity (7.9 mg cell l(-1) h(-1)), biomass yield (0.041 g cell g(-1) sugar) and specific growth rate (0.066 h(-1)). CONCLUSIONS: The limitations to the growth of O. oeni were pH and inhibition by end product resulting in poor utilization of the medium with low cell yields. The cell productivity of the system can be improved by the appropriate use of mixed sugar growth medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study uniquely showed that appropriate sugar mixtures with the correct environmental conditions can significantly improve the productivity of O. oeni cultures.     

Role of calonyctin on free sugars in relation to starch accumulation in developing sweet potatoes

Calonyctin, a natural plant growth regulator extracted from the leaves of Calonyction aculeatum (L.) House, can promote crop growth and increase crop yield. The specific reasons for this response are unknown. This study was conducted to determine the effect of calonyctin treatment on the free sugars of sweet potato [Ipomoea batatas (L.) Lam.] as related to starch accumulation. The sweet potatoes were grown in the field in 1992, treated by foliar spray with Calonyctin concentrations of 0 (control) and 0.1 activity unit (CTSP) at 20 days after planting (DAP) at the rate of 190 liters of diluted solution/ha., and sampled periodically to determine free sugars. The response of sweet potato to calonyctin was first detected at 40 days after treatment (on 60 DAP). Data indicated that calonyctin treatment significantly increased starch synthesis in storage roots, decreased the fluctuation tendency of total sugar level during the growth period, and kept the sugar level relatively constant with a gradual rise regardless of variations in weather. The level of the reducing sugars in CTSP leaves was higher at 60 and 160 DAP and lower at 100, 120, and 140 DAP. During rainy days (100 DAP), the reducing sugars in CTSP storage roots remained at a lower level when those in controls reached high levels. The sucrose content in CTSP leaves was 40–138% greater than that in controls except at 80 and 120 DAP, and the ratio of sucrose to total nonreducing sugars remained at 100% in CTSP leaves even on rainy and cool days and above 96% in CTSP storage roots except on cool days (140 and 160 DAP), suggesting that calonyctin treatment promoted the synthesis and transfer of sucrose and supplied abundant sugar precursors for starch accumulation in storage roots.Abbreviations DAP days after planting - CTSP calonyctin-treated sweet potato with 0.1 activity unit     

Studies of the Utilization of Coconut Water Waste for the Production of the Food Yeast Saccharomyces fragilis

The accepted food yeast Saccharomyces fragilis was grown in batch and chemostat culture on coconut water and on a simulated coconut-water medium containing glucose, fructose, sucrose and sorbitol, to provide kinetic data for a feasibility study of microbial protein production. Analyses of growth on individual and mixed carbon substrates were made to determine sugar assimilation patterns in batch and chemostat cultures on coconut water. Growth on the polyol produced a much reduced specific growth rate, assimilation rate, growth yield and productivity compared to growth on the sugars. In mixed substrate fermentations a sequential utilization of the carbohydrates occurred. Both the monosaccharides repressed invertase synthesis and all three sugars repressed sorbitol assimilation. Complete carbon assimilation was only obtained by prolonged batch fermentation or in chemostat cultures at low dilution rates (<0.10 h^-1). Supplementation of coconut water with biotin and nicotinic acid increased biomass yields in chemostat cultures.     

Effect of aluminium on metabolism of starch and sugars in growing rice seedlings

The effect of increasing concentrations of Al₂(SO₄)₃ in situ on the content of starch, sugars and activity behaviour of enzymes related to their metabolism were studied in growing seedlings of two rice cvs. Malviya-36 and Pant-12 in sand cultures. Al₂(SO₄)₃ levels of 80 and 160 μM in the growth medium caused an increase in the contents of starch, total sugars as well as reducing sugars in roots as well as shoots of the rice seedlings during a 5–20 days growth period. The activities of the enzymes of starch hydrolysis α-amylase, β-amylase and starch phosphorylase declined in Al-exposed seedlings, whereas the activities of sucrose hydrolyzing enzymes sucrose synthase and acid invertase increased in the seedlings due to Al³⁺ treatment. The enzyme of sucrose synthesis, sucrose phosphate synthase showed decreased activity in Al³⁺ treated seedlings compared to controls. Results suggest that Al³⁺ toxicity in rice seedlings impairs the metabolism of starch and sugars and favours the accumulation of hexoses by enhancing the activities of sucrose hydrolyzing enzymes.     

Sensitivity of a Bacteroides melaninogenicus strain to monosaccharides: effect on enzyme induction.

The inhibition of growth in Bacteroides melaninogenicus by sugars in described. Monosaccharides such as D-glucose, D-galactose, D-mannose, and D-fructose are inhibitory at low concentrations, whereas the disaccharides sucrose and lactose are not inhibitory even at high concentrations. The major inhibitory effect of the sugar is found during the transition of lag to logarithmic growth phases. There was no primary effect of D-glucose on protein, ribonucleic acid, or deoxyribonucleic acid synthesis on cells in transition from lag to logarithmic growth. However, the addition of glucose or galactose completely abolished the induction of 3-ketodihydrosphingosine synthetase by vitamin K in vitamin K-depleted cells. Futhermore, in cells which were not vitamin K depleted, the level of this enzyme was drastically reduced by the addition of the sugar. Cyclic adenosine 5-monophosphate was unable to reverse the growth inhibition produced by glucose. In actively growing cultures, addition of sugar slows the growth rate. In these experiments the level of 3-ketodihydrosphingosine synthetase fell only after the cells had assumed the slower rate of growth. There were two indications that D-galactose was more inhibitory than D-glucose; in the presence of 0.1% D-galactose cells in lag phase did not show the increase in turbidity found in similar cells placed in medium with 0.1% D-glucose, and also D-galactose caused a greater decrease in the growth rate of actively growing cultures than was found with D-glucose. These studies suggest that the inhibitory effect of monosaccharides in lag leads to logarithmic growth transition can be ascribed to an effect on enzyme induction. On the other hand, the ability of many monosaccharides to inhibit growth, and the greater inhibitory property of D-galactose compared with D-glucose, suggests that other mechanisms may be operative as well.     

CLONAL VARIATION OF GRAPE-STEM AND PHYLLOXERA-GALL CALLUS GROWING IN VITRO IN DIFFERENT CONCENTRATIONS OF SUGARS

Arya , H. C., A. C. Hildebrandt , and A. J. Riker . (University of Wisconsin, Madison, Wisconsin.) Clonal variation of grape-stem and Phylloxera-gall callus growing in vitro in different concentrations of sugars. Amer. Jour. Bot. 49(4): 368–372. Illus. 1962.—The original callus grown from normal tissue and that grown from gall tissue contained mixtures of different kinds of cells. To study the variability, a large number of clones were developed by single-cell technique. From these, 6 clones were selected for detailed study. Growth was compared of 6 single-cell clones established in vitro; 3 from normal grape stem and 3 from gall tissues incited by Phylloxera vastatrix Planch. The clones were stable in growth rate (fast, medium, and slow) when grown on modified White's basal medium supplemented with coconut milk, α-naphthaleneacetic acid (NAA), and calcium pantothenate. Growth was measured after 6 weeks as the average wet weight on concentrations from 0.06–4.0% of sucrose, d(+) dextrose, d(-) levulose, d(+) mannose, d(+) galactose, and d(+) lactose, respectively. Every sugar, except mannose, was a suitable source of carbon. The cells were not all alike in their growth response to different sugars. Gall- and normal-tissue clones grew best with 0.125% sucrose, levulose, and galactose. With dextrose and lactose, optimum yields were obtained at the 1.0% sugar level. Growth of fast-, medium-, and slow-growing clones was altered with the type of sugar. Gall and normal tissues were differentiated from each other when grown on mannose in which gall tissues grew best at 0.125% and normal at 1.0% levels. Gall tissues as a group were able to grow better with mannose than the corresponding normal ones. Levulose, on the other hand, favored growth of normal clones in comparison to that of diseased ones. Sugars varied in their inhibitory influence at the 4.0% level. Dextrose, levulose, and lactose at 1.0% proved better than sucrose for the growth of all except one fast-growing gall clone which grew best with sucrose. However, at 0.125% sugar levels, even in cases where high yields were obtained, the physical character of the tissues changed to dry, brownish, and very friable. Thus, the original callus from normal and gall tissues contained cells with diverse characteristics. The various clones developed used the same sugars but varied strikingly in the rate and type of growth on certain sugars.     

Cucumber mosaic virus infection affects sugar transport in melon plants

Viral infection often affects carbon assimilation and metabolism in host plants. To better understand the effect of cucumber mosaic virus (CMV) infection on sugar transport, carbohydrate levels and the amounts of the various sugars in the phloem sap were determined in infected melon (Cucumis melo L.) plants. Source leaves infected with CMV were characterized by high concentrations of reducing sugars and relatively low starch levels. The altered level of carbohydrates was accompanied by increased respiration and decreased net photosynthetic rates in the infected leaves. Although stachyose was the predominant sugar in phloem sap collected from petioles of control leaves, sucrose (Suc) was a major sugar in the phloem sap of infected leaves. Moreover, analyses of the newly fixed (14)CO(2) revealed a high proportion of radioactive Suc in the phloem sap of infected leaves 60 min post-labeling. The alteration in phloem sap sugar composition was found in source, but not old, leaves. Moreover, elevations in Suc concentration were also evident in source leaves that did not exhibit symptoms or contain detectable amounts of virus particles. The mode by which CMV infection may cause alterations in sugar transport is discussed in terms of the mechanism by which sugars are loaded into the phloem of cucurbit plants.     

Sucrose and starch metabolism during Fargesia yunnanensis shoot growth

Bamboo is one of the fastest growing plants in the world, but their shoot buds develop very slowly. Information about the sugar storage and metabolism during the shoot growth is lacking. In the present study, we determined the activity of sucrose and starch metabolizing enzymes during the developmental period of Fargesia yunnanensis from shoot buds to the young culms that have achieved their full height. The soluble sugars and starch contents were also determined and analyzed in shoot buds and shoots at different developmental stages. The results showed that there were higher sucrose contents in shoot buds than shoots, which coincides with the sweeter taste of shoot buds. As the shoot buds sprouted out of the ground, the starch and sucrose were depleted sharply. Coupled with this, the activity of soluble acid invertase (SAI), cell wall-bound invertase (CWI), sucrose synthase at cleavage direction (SUSYC) and starch phosphorylase (STP) increased significantly in the rapidly elongating internodes. These enzymes dominated the rapid elongation of internodes. The activities of SAI, CWI, SUSYC and STP and adenosine diphosphate-glucose pyrophosphorylase were higher as compared to other enzymes in the shoot buds, but were far lower than those in the developing shoots. The slow growth of shoot buds was correlated with the low activity of these enzymes. These results complement our understanding of the physiological differences between shoot buds and elongating shoots and ascertain the physiological mechanism for the rapid growth of bamboo shoots.