Direct current electrical field effects on intact plant organs.

Intact plant tissues (of hypocotyls, radicles, cotyledons and leaves) were contracted by applying a low DC electrical field through them. Stomatal opening as a result of the electrical treatment of leaves was observed, presumably due to the differential influence of the electrical treatment on guard cell turgor pressure versus turgor pressure of the surrounding epidermal cells. In addition, leakage of minerals from the treated leaves was detected (higher contents of potassium, sodium, calcium and sulfur), as was leakage of betanin from electrically treated red beet roots (higher OD value of the immersion solution with increasing time of applied electrical field). Application of such a treatment can be used for initial drying or as part of another more drastic drying method. The advantages of this method lie in its nonthermal character and its potential to increase the quality of processed foods by maintaining their "like-fresh" quality, e.g., fruits and vegetables that are less damaged by browning. An understanding of the mechanism involved in this nonthermal application can help in controlling the process and predicting its outcome.

Enhanced bud and bulblet regeneration from bulbs of Nerine sarniensis cultured in vitro.

Nerine (Nerine sarniensis) cv. Salmon Supreme in vitro-grown bulblets, 7-9 mm in diameter, were cut in half longitudinally and used for adventitious bud initiation following dissection of the roots and two-thirds of the upper part of the bulblets. The terminal apex was injured with a hot, sterile microscope dissecting needle. The highest number of buds formed (seven to nine buds per halved bulblet) on a semi-solid Murashige and Skoog (MS) basal salts medium supplemented with 3% sucrose and either 1 microM 6-benzylaminopurine (BA) and 1 microM alpha-naphthaleneacetic acid (NAA) or 0.5 microM BA and 0.1 microM NAA. Bulblet halves were cultured in the dark for 11-13 weeks with one subculture after 6 weeks. Anatomical studies indicated that the initiation of adventitious buds on the abaxial side of the inner scales of the halved bulblet was adjacent to the basal plate and started from the leaf primordium and a meristematic bulge. Buds developed directly into small bulblets after they were transferred to semi-solid MS basal salts medium supplemented with 6% sucrose, 10 microM indole-3-butyric acid (IBA) and 0.25% activated charcoal. Small bulblets cultured in liquid MS medium supplemented with additional KH(2)PO(4 )(170 mg l(-1)), 6% sucrose and 0.1 microM NAA under a 16/8-h (light/dark) photoperiod for 8 weeks grew into larger bulbs faster than those cultured on semi-solid medium. The bulbs were rooted on a semi-solid medium after 4 weeks and then transferred to the soil. As many as 18 bulblets developed and rooted from one in vitro-grown bulb after 25-27 weeks.

The Role of Autumn Infections in the Progression of Fire Blight Symptoms in Perennial Pear Branches.

The role of autumn infections in the progression of fire blight (caused by Erwinia amylovora) symptoms in perennial pear branches was studied in orchard-grown trees in Israel. The extent of symptom progression and the final length of fire blight cankers in perennial branches were variably affected by the vigor of the trees and the season of infection. Following spring infections, when all trees supported active shoot growth, fire blight symptoms progressed more rapidly and to longer distances in trees that exhibited high vigor (i.e., with numerous annual shoots on most terminal branches) than in low-vigor trees (i.e., few or no annual shoots on terminal branches). Irrespective of the vigor of the trees, the progression of fire blight symptoms in perennial branches ceased between mid-May and mid-July, and only a small proportion (0 to 14.2%) of the infections had invaded main limbs or trunks of trees. Progression of fire blight symptoms following autumn infections was related to the preceding summer (August to No-vember) shoot regrowth: in trees in which the shoots did not restore their growth in the summer, the rate of symptom progression in perennial branches was higher in trees with a low vigor than in those with a high vigor, whereas for those with summer regrowth the relationship between rates of symptom expression was reversed. Irrespective of the vigor group and of whether there was summer regrowth, symptoms in perennial branches continued to progress through the winter until the following spring. Most of the autumn infections (50 to 78.5%) that developed in susceptible trees had invaded main limbs or trunks of trees. The results of this study indicate that factors related to host phenology and physiology, rather than factors related to environmental influences (such as temperature), govern the extent, rate, and duration of fire blight progression in perennial pear branches. Furthermore, it turned out that autumn infections play a substantial role in fire blight epidemiology in Israel.

Analysis of celery (Apium graveolens) mannitol dehydrogenase (Mtd) promoter regulation in Arabidopsis suggests roles for MTD in key environmental and metabolic responses.

Of the growing list of promising genes for plant improvement, some of the most versatile appear to be those involved in sugar alcohol metabolism. Mannitol, one of the best characterized sugar alcohols, is a significant photosynthetic product in many higher plants. The roles of mannitol as both a metabolite and an osmoprotectant in celery (Apium graveolens) are well documented. However, there is growing evidence that 'metabolites' can also have key roles in other environmental and developmental responses in plants. For instance, in addition to its other properties, mannitol is an antioxidant and may have significant roles in plant-pathogen interactions. The mannitol catabolic enzyme mannitol dehydrogenase (MTD) is a prime modulator of mannitol accumulation in plants. Because the complex regulation of MTD is central to the balanced integration of mannitol metabolism in celery, its study is crucial in clarifying the physiological role(s) of mannitol metabolism in environmental and metabolic responses. In this study we used transformed Arabidopsis to analyze the multiple environmental and metabolic responses of the Mtd promoter. Our data show that all previously described changes in Mtd RNA accumulation in celery cells mirrored changes in Mtd transcription in Arabidopsis. These include up-regulation by salicylic acid, hexokinase-mediated sugar down-regulation, and down-regulation by salt, osmotic stress and ABA. In contrast, the massive up-regulation of Mtd expression in the vascular tissues of salt-stressed Arabidopsis roots suggests a possible role for MTD in mannitol translocation and unloading and its interrelation with sugar metabolism.

Phloem-Specific Expression of the Tobacco Mosaic Virus Movement Protein Alters Carbon Metabolism and Partitioning in Transgenic Potato Plants.

The tobacco mosaic virus movement protein (TMV-MP) has pleiotropic effects when expressed in transgenic tobacco (Nicotiana tabacum) plants. In addition to its ability to increase the plasmodesmal size-exclusion limit, the TMV-MP alters carbohydrate metabolism in source leaves and dry matter partitioning between the various plant organs. In the present study the TMV-MP was expressed under the control of a phloem-specific promoter (rolC), and this system was employed to further explore the potential sites at which the TMV-MP exerts its influence over carbon metabolism and transport in transgenic potato (Solanum tuberosum) plants. Immunohistochemical analyses indicated that the TMV-MP was localized mainly to phloem parenchyma and companion cells. Starch and sucrose accumulated in source leaves of these plants to significantly higher levels compared with control potato lines. In addition, the rate of sucrose efflux from excised petioles was lower compared with control plants. Furthermore, under short-day conditions, carbon partitioning was lower to the roots and higher to tubers in rolC plants compared with controls. These results are discussed in terms of the mode(s) by which the TMV-MP exerts its influence over carbon metabolism and photoassimilate translocation.

Subcellular localization of celery mannitol dehydrogenase. A cytosolic metabolic enzyme in nuclei.

Mannitol dehydrogenase (MTD) is the first enzyme in mannitol catabolism in celery (Apium graveolens L. var dulce [Mill] Pers. cv Florida 638). Mannitol is an important photoassimilate, as well as providing plants with resistance to salt and osmotic stress. Previous work has shown that expression of the celery Mtd gene is regulated by many factors, such as hexose sugars, salt and osmotic stress, and salicylic acid. Furthermore, MTD is present in cells of sink organs, phloem cells, and mannitol-grown suspension cultures. Immunogold localization and biochemical analyses presented here demonstrate that celery MTD is localized in the cytosol and nuclei. Although the cellular density of MTD varies among different cell types, densities of nuclear and cytosolic MTD in a given cell are approximately equal. Biochemical analyses of nuclear extracts from mannitol-grown cultured cells confirmed that the nuclear-localized MTD is enzymatically active. The function(s) of nuclear-localized MTD is unknown.

Immunolocalization of mannitol dehydrogenase in celery plants and cells.

Immunolocalization of mannitol dehydrogenase (MTD) in celery (Apium graveolens L.) suspension cells and plants showed that MTD is a cytoplasmic enzyme. MTD was found in the meristems of celery root apices, in young expanding leaves, in the vascular cambium, and in the phloem, including sieve-element/companion cell complexes, parenchyma, and in the exuding phloem sap of cut petioles. Suspension cells that were grown in medium with mannitol as the sole carbon source showed a high anti-MTD cross-reaction in the cytoplasm, whereas cells that were grown in sucrose-containing medium showed little or no cross-reaction. Gel-blot analysis of proteins from vascular and nonvascular tissues of mature celery petioles showed a strong anti-MTD sera cross-reactive band, corresponding to the 40-kD molecular mass of MTD in vascular extracts, but no cross-reactive bands in nonvascular extracts. The distribution pattern of MTD within celery plants and in cell cultures that were grown on different carbon sources is consistent with the hypothesis that the Mtd gene may be regulated by sugar repression. Additionally, a developmental component may regulate the distribution of MTD within celery plants.

Turgor regulation of sucrose transport in sugar beet taproot tissue.

Sink tissues that store osmotically active compounds must osmoregulate to prevent excessively high turgor. The ability to regulate turgor may be related to membrane transport of solutes and thus sink strength. To study this possibility, the kinetics of sugar uptake were determined in sugar beet (Beta vulgaris L.) taproot tissue discs over a range of cell turgors. Sucrose uptake followed biphasic kinetics with a high affinity saturating component below 20 millimolar and a low affinity linear component at higher concentrations. Glucose uptake exhibited only simple saturation type kinetics. The high affinity saturating component of sucrose and glucose uptake was inhibited by increasing cell turgor (decreasing external mannitol concentrations). The inhibition was evident as a decrease in V(max) but no effect on K(m). Sucrose uptake by tissue equilibrated in dilute buffer exhibited no saturating component. Ethylene glycol, a permeant osmoticum, had no effect on uptake kinetics, suggesting that the effect was due to changes in cell turgor and not due to decreased water potential per se. p-(Chloromercuri)benzene sulfonic acid (PCMBS) inhibited sucrose uptake at low but not high cell turgor. High cell turgor caused the tissue to become generally leaky to potassium, sucrose, amino acids, and reducing sugars. PCMBS had no effect on sucrose leakage, an indication that the turgor-induced leakage of sucrose was not via back flow through the carrier. The ability of the tissue to acidify the external media was turgor dependent with an optimum at 300 kilopascals. Acidification was sharply reduced at cell turgors above or below the optimum. The results suggest that the secondary transport of sucrose is reduced at high turgor as a result of inhibition of the plasma membrane ATPase. This inhibition of ATPase activity would explain the reduced V(max) and leakiness to low molecular weight solutes. Cell turgor is an important regulator of sucrose uptake in this tissue and thus may be an important determinant of sink strength in tissues that store sucrose.

Stereospecificity of the glucose carrier in sugar beet suspension cells.

The stereospecificity of the binding site on the glucose carrier system in sugar beet suspension culture cells was determined using a series of aldo and keto hexose sugars and sugar alcohols. Specificity was determined as competition with [(14)C]glucose transport and glucose/proton symport.The binding site of the glucose carrier system was specific for the stereo orientation of the three equatorial OH groups on the three carbons opposite the oxygen and for the CH(2)OH group. Hexopyranose isomers with the same orientation at the three OH groups (carbons 2, 3, and 4 of C-1 d-glucose), but not with the CH(2)OH group, have only little (1-C d-glucose) or no effect (1-C d-idose and myoinositol) on d-glucose uptake. The C-1 l-sorbose molecule matches the C-1 d-glucose at many points including the stereo configuration of the CH(2)OH group, but it had no effect on d-glucose uptake perhaps because of an interference of the OH group adjacent to the CH(2)OH substituent. The d-glucose analogs, 3-O-methylglucose and glucosamine, were the most effective in binding to the glucose carrier. The isomers d-fructose, d-galactose, and d-mannose have separate distinctive proton cotransport systems. However, in starved cells they compete with d-glucose uptake, but the competition is for the available energy and not the carrier binding site.