Donald L. Hendrix

Increased Accumulation of Carbohydrates and Decreased Photosynthetic Gene Transcript Levels in Wheat Grown at an Elevated CO2 Concentration in the Field

Repression of photosynthetic genes by increased soluble carbohydrate concentrations may explain acclimation of photosynthesis to elevated CO2 concentration. This hypothesis was examined in a field crop of spring wheat (Triticum aestivum L.) grown at both ambient (approximately 360 [mu]mol mol-1) and elevated (550 [mu]mol mol-1) atmospheric CO2 concentrations using free-air CO2 enrichment at Maricopa, Arizona. The correspondence of steady-state levels of mRNA transcripts (coding for the 83-kD photosystem I apoprotein, sedoheptulose-1,7-bisphosphatase, phosphoribulokinase, phosphoglycerokinase, and the large and small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase) with leaf carbohydrate concentrations (glucose-6-phosphate, glucose, fructose, sucrose, fructans, and starch) was examined at different stages of crop and leaf development and through the diurnal cycle. Overall only a weak correspondence between increased soluble carbohydrate concentrations and decreased levels for nuclear gene transcripts was found. The difference in soluble carbohydrate concentration between leaves grown at elevated and current ambient CO2 concentrations diminished with crop development, whereas the difference in transcript levels increased. In the flag leaf, soluble carbohydrate concentrations declined markedly with the onset of grain filling; yet transcript levels also declined. The results suggest that, whereas the hypothesis may hold well in model laboratory systems, many other factors modified its significance in this field wheat crop.

Photosynthesis of cotton plants exposed to elevated levels of carbon dioxide in the field

The cotton (Gossypium hirsutum L.) plant responds to a doubling of atmospheric CO2 with almost doubled yield. Gas exchange of leaves was monitored to discover the photosynthetic basis of this large response. Plants were grown in the field in open-top chambers with ambient (nominally 350 μl/l) or enriched (nominally either 500 or 650 μl/l) concentrations of atmospheric CO2. During most of the season, in fully-irrigated plants the relationship between assimilation (A) and intercellular CO2 concentration (ci) was almost linear over an extremely wide range of ci. CO2 enrichment did not alter this relationship or diminish photosynthetic capacity (despite accumulation of starch to very high levels) until very late in the season, when temperature was somewhat lower than at midseason. Stomatal conductance at midseason was very high and insensitive to CO2, leading to estimates of ci above 85% of atmospheric CO2 concentration in both ambient and enriched chambers. Water stress caused A to show a saturation response with respect to ci, and it increased stomatal closure in response to CO2 enrichment. In fully-irrigated plants CO2 enrichment to 650 μl/l increased A more than 70%, but in water-stressed plants enrichment increased A only about 52%. The non-saturating response of A to ci, the failure of CO2 enrichment to decrease photosynthetic capacity for most of the season, and the ability of the leaves to maintain very high ci, form in part the basis for the very large response to CO2 enrichment.

A thermoprotective role for sorbitol in the silverleaf whitefly, Bemisia argentifolii.

Accumulation of polyols in insects is well known as a cold-hardening response related to overwintering or to protection against cold shock. The silverleaf whitefly (Bemisia argentifolii, Bellows and Perring) is a major insect pest in tropical and subtropical regions where heat stress and desiccation pose formidable threats to survival. We found that sorbitol levels increased ten-fold when whiteflies were exposed to elevated temperatures. Sorbitol levels rose from 0.16nmolwhitefly(-1) at 25 degrees C to 1.59nmolwhitefly(-1) at 42 degrees C. Sorbitol levels fluctuated diurnally under glasshouse and field conditions increasing ten-fold from morning to early afternoon. Feeding experiments on artificial diets showed that both temperature and dietary sucrose concentration were key factors influencing sorbitol accumulation. Cell free extracts prepared from adult whiteflies catalyzed NADPH-dependent fructose reduction, but were unable to reduce glucose with either NADPH or NADH. Radiotracer experiments with labeled glucose and fructose showed that fructose was the immediate precursor of sorbitol. Thus, sorbitol synthesis in the whitefly is apparently unconventional, involving conversion of fructose by a novel NADPH-dependent ketose reductase. We propose that sorbitol accumulation is a mechanism for thermoprotection and osmoregulation in the silverleaf whitefly, allowing the insect to thrive in environments conducive to thermal and osmotic stress.

Bemisiose: an unusual trisaccharide in Bemisia honeydew☆

A gradient high performance anion-exchange chromatography analysis of Bemisia honeydew was carried out in order to analyze the sugar mixture more completely. From the results of the experiments by using chemical-ionization and electron-ionization gas chromatography, enzyme digestion, and H and C NMR that showed this saccharide to be a trisaccharide consisting entirely of a alpha-D-glucose residues, a structure was proposed. The common name bemisiose is suggested for this sugar because it was found in Bemisia honeydew.

Sucrose Synthase Localization during Initiation of Seed Development and Trichome Differentiation in Cotton Ovules

Sucrose synthase in cotton (Gossypium hirsutum L.) ovules was immunolocalized to clarify the relationship between this enzyme and (a) sucrose import/utilization during initiation of seed development, (b) trichome differentiation, and (c) cell-wall biosynthesis in these rapidly elongating “fibers.” Analyses focused on the period immediately before and after trichome initiation (at pollination). Internal tissues most heavily immunolabeled were the developing nucellus, adjacent integument (inner surface), and the vascular region. Little sucrose synthase was associated with the outermost epidermis on the day preceding pollination. However, 1 d later, immunolabel appeared specifically in those epidermal cells at the earliest visible phase of trichome differentiation. The day following pollination, these cells had elongated 3- to 5-fold and showed a further enhancement of sucrose synthase immunolabel. Levels of sucrose synthase mRNA also increased during this period, regardless of whether pollination per se had occurred. Timing of onset for the cell-specific localization of sucrose synthase in young seeds and trichome initials indicates a close association between this enzyme and sucrose import at a cellular level, as well as a potentially integral role in cell-wall biosynthesis.

Polyol metabolism in homopterans at high temperatures: accumulation of mannitol in aphids (Aphididae: Homoptera) and sorbitol in whiteflies (Aleyrodidae: Homoptera)

Examination of four species of aphid and four of whitefly showed that mannitol was present in each species of aphid, while sorbitol was present in the whitefly species. In the cotton aphid, Aphis gossypii Glover, the total body content of mannitol was considerably higher at noon than during the early morning. A similar increase in the sorbitol content of the silverleaf whitefly, Bemisia argentifolii Bellows and Perring, was also demonstrated. In both species, polyol synthesis is stimulated by elevated temperatures. Enzyme assays were used to show that fructose is the substrate for mannitol synthesis in A. gossypii. The enzyme catalyzing this reaction, an NADP(H)-dependent ketose reductase/mannitol dehydrogenase, is analogous to the NADP(H)-dependent ketose reductase/sorbitol dehydrogenase that produces sorbitol in whiteflies. Western blot analysis verified that A. gossypii does not contain a protein that cross-reacts with antibodies against B. argentifolli NADP(H)-dependent ketose reductase/sorbitol dehydrogenase, whereas the greenhouse whitefly, Trialeurodes vaporariorum Westwood, does. Analysis of sugars in honeydew from aphids and whiteflies showed that the sugars in the excrement from these insects are very different from the sugars present in their bodies. Only small amounts of mannitol and sorbitol are excreted in the honeydew from these insects. Sorbitol accumulation provides a mechanism for thermo- and osmoprotection in whiteflies. Mannitol appears to function in a similar capacity in aphids.

Effect of sucrose concentration on carbohydrate metabolism in Bemisia argentifolii: Biochemical mechanism and physiological role for trehalulose synthesis in the silverleaf whitefly

Uptake and metabolism of sucrose by adult silverleaf whiteflies (Bemisia argentifolii) were investigated on defined diets containing sucrose concentrations from 3 to 30% (wv). At an optimal pH of 7, the volume of liquid ingested decreased with increasing dietary sucrose concentration, but the amount of sucrose ingested showed a net increase. Above a dietary sucrose concentration of about 10%, a greater amount of the ingested carbon was excreted by the whiteflies than was retained, and the proportion that was excreted increased progressively with increasing dietary sucrose concentration. Carbohydrate analysis showed that the composition of excreted honeydew changed from predominantly glucose and fructose at low dietary sucrose concentrations to predominantly trehalulose at high concentrations, with little change in the proportion of larger oligosaccharides. Measurements of whitefly trehalulose synthase and sucrase activities revealed that the enzymatic potential for metabolizing sucrose shifted from favoring sucrose hydrolysis at low sucrose concentrations to sucrose isomerization at high sucrose concentrations. Thus, the amount of trehalulose synthesized by the silverleaf whitefly was directly related to the properties of trehalulose synthase and sucrase and the concentration of sucrose in the diet. We propose that trehalulose is synthesized for excretion when the carbon input from sucrose is in excess of metabolic needs.

The apoplastic pool of abscisic acid in cotton leaves in relation to stomatal closure

Suboptimal nitrogen nutrition, leaf aging, and prior exposure to water stress all increased stomatal closure in excised cotton (Gossypium hirsutum L.) leaves supplied abscisic acid (ABA) through the transpiration stream. The effects of water stress and N stress were partially reversed by simultaneous application of kinetin (N6-furfurylaminopurine) with the ABA, but the effect of leaf aging was not. These enhanced responses to ABA could have resulted either from altered rates of ABA release from symplast to apoplast, or from some “post-release” effect involving ABA transport to, or detection by, the guard cells. Excised leaves were preloaded with [14C]ABA and subjected to overpressures in a pressure chamber to isolate apoplastic solutes in the exudate. Small quantities of 14C were released into the exudate, with the amount increasing greatly with increasing pressure. Over the range of pressures from 1 to 2.5 MPa, ABA in the exudate contained about 70% of the total 14C, and a compound co-chromatographing with phaseic acid contained over half of the remainder. At a low balancing pressure (1 MPa), release of 14C into the exudate was increased by N stress, prior water stress, and leaf aging. Kinetin did not affect 14C release in leaves of any age, N status, or water status. Distribution of ABA between pools can account in part for the effects of water stress, N stress, and leaf age on stomatal behavior, but in the cases of water stress and N stress there are additional kinetinreversible effects, presumably at the guard cells.

FEEDING RATES AND CARBOHYDRATE METABOLISM BY BEMISIA TABACI (HOMOPTERA : ALEYRODIDAE) ON DIFFERENT QUALITY PHLOEM SAPS

Abstract.The effects of water stress on phloem sap quality of the melon, Cucumis melo, and how this, in turn, has an impact on the sweet potato whitefly, Bemisia tabaci were studied. Melon plants were grown under watering regimes that produced plants with or without water stress. Plants showed strong developmental responses to the treatments; water‐stressed plants were shorter, with fewer, smaller leaves than those without stress. There was, however, no effect of plant water stress on the development period of whiteflies feeding on these plants, or on the weights of male or female adults. Honeydew production was used as an indirect measure to test whether the absence of insect developmental or behavioural effects was due to differential phloem sap ingestion. Feeding rates on the stressed plants were almost half those on unstressed plants, and there was also variation in the daily pattern of honeydew production. Phloem sap and honeydew were analysed to determine why the feeding behaviours differed. Amino acid composition of the phloem sap was similar in both groups of plants, but carbohydrate concentrations were greater in water‐stressed plants, indicating that lower feeding rates may have been due either to the physical or nutritional quality of the phloem sap. The honeydew of insects that were feeding on water‐stressed plants contained a greater concentration of carbohydrate than those on unstressed plants, and was composed of a significantly greater proportion of glucose and the disaccharide, trehalulose. This isomerization of more complex sugars from those in the diet suggests that B. tabaci uses a mechanism of osmoregulation to actively maintain its internal water status. It is concluded that transient conditions of water stress in this host plant do not affect the development of B. tabaci, due to physiological and behavioural changes in response to diets with different nutritional and physical properties. The implications of this finding for the feeding biology of B. tabaci on desert‐grown crops are discussed.

EFFECT OF HIGH TEMPERATURE ON THE METABOLIC PROCESSES AFFECTING SORBITOL SYNTHESIS IN THE SILVERLEAF WHITEFLY, BEMISIA ARGENTIFOLII

Whiteflies accumulate the polyhydric alcohol, sorbitol, when exposed to temperatures greater than about 30 degrees C. Feeding experiments using artificial diets containing labeled sucrose showed that more of the label was incorporated into whitefly bodies and less was excreted in the honeydew when feeding was conducted at 41 compared with 25 degrees C. Analysis of the components of the honeydew showed that more of the excreted label was in glucose and fructose and less in trehalulose at 41 degrees C than at 25 degrees C. A similar effect of temperature on honeydew composition occurred for whiteflies feeding on cotton leaves. Measurement of the activities of glycolytic, pentose-phosphate and polyol pathway enzymes at 30 and 42 degrees C showed that NADPH-dependent ketose reductase/sorbitol dehydrogenase (NADPH-KR/SDH), sucrase, glucokinase and glucose-6-phosphate dehydrogenase activities were stimulated to a greater extent at 42 degrees C than trehalulose synthase and fructokinase. NAD(+)-sorbitol dehydrogenase (NAD(+)-SDH) activity was inhibited at 42 degrees C. We propose that high temperature alters metabolic activity in a way that increases the availability of fructose and stimulates pentose-phosphate pathway activity, providing both the substrate and coenzyme for sorbitol synthesis. High temperature also increases the activity of NADPH-KR/SDH, the enzyme in whiteflies that synthesizes sorbitol, but inhibits the activity of NAD(+)-SDH, the enzyme that degrades sorbitol.