光合同化物韧皮部卸载途径和机制

介绍近年来同化物在库器官中的卸载途径、机制和研究方法的研究进展,并简略讨论了该领域研究所面临的问题。     

Phloem Unloading in Developing Leaves of Sugar Beet : II. Termination of Phloem Unloading

Phloem unloading in developing leaves of Beta vulgaris L. (`Klein E' multigerm) occurred from successively higher order branches of veins as leaves matured. Phloem unloading was studied in autoradiographs of leaf samples taken at various times during the arrival of a pulse of ¹⁴C-labeled photoassimilate. Extension of mass flow of sieve element contents into leaf vein branches was determined from the high level of radiolabel in veins soon after first arrival of the pulse. Rapid entry, indicative of mass flow through open sieve pores, occurred down to the fourth division of veins in young, importing leaves and to the fifth or terminal branch in importing regions near the zone of transition from sink to source. The rate of unloading decreased with leaf age, as evidenced by the increased time required for the vein-mesophyll demarcation to become obscured. The rate of import per unit leaf area, measured by steady state labeling with ¹⁴CO₂ also decreased as a leaf matured. The decline in import appeared to result from progressive changes that increased resistance to unloading of sieve elements and eventually terminated phloem unloading.     

Apoplastic Phloem Unloading in the Stem of Bean

Sucrose has been found in the apoplast of bean stems at a concentrationof 25–60 mM with an axial concentration gradient in theappropriate direction for Munch translocation. Removal of theepidermis from a 50 mm length of stem enabled the washout oflabelled photosynthate from the apoplast. The rate of labelwashout was strongly dependent on temperature, and the rateincreased on blockage of phloem pathways to the main sink forthat assimilate. Washout did not reduce when the bathed tissuewas plasmolyzed. We propose that sucrose is unloaded from thephloem into the apoplast, and a sucrose concentration is maintainedthere by a balance of sucrose uptake into sink tissue or reloadinginto the phloem. It is proposed that the apoplastic pool ofphotosynthate can act to buffer sudden changes in phloem contentswhen there are rapid changes in source-sink configuration. Key words: Sucrose, Phaseolus vulgaris, Apoplast, Phloem unloading     

Xylem, Phloem and Transpiration Flows in Developing Apple Fruits

Xylem, phloem and transpiration flows were measured in developingfruits of the apples Royal Gala and Cox's Orange Pippin at early,mid and late stages during their development. Fruit volume growthwas first computed from measurements of diameter made usinga system of sensitive displacement transducers. Xylem, phloemand transpiration flow components (of which fruit volume growthis the integral) were then separated using a scheme of treatmentswhich disabled one or other flow at a time. Changes observed during development in the patterns of the threeflows are in general agreement with expectations based on lessdirect observations (loc. cit. Ferguson and Watkins, 1989).Recognizing the distinctive mineral compositions of xylem andphloem streams, the changes also accommodate, and seem to explain,published observations of mineral accumulation in apple. Significant differences in the pattern of xylem and phloem flowwere observed between the varieties examined. These may explainvarietal differences in susceptibility to the mineral imbalancedisorder bitter-pit. Xylem flows were shown to reverse at times; that is they flowedfrom fruit to tree. This occurred particularly during periodsof high evaporative demand. Xylem reversal is of importanceto the overall water economy of a tree but may also have specialimportance to the mineral composition of the fruit. Key words: Mineral nutrition, bitter-pit, apple, xylem, phloem, transpiration, water balance     

韧皮部运输和防御作用的分子机理

韧皮部是维管系统的重要组成部分,在物质运输、信息传递、机械支持以及防御作用中发挥重要作用。近年来,在建立了一系列研究方法的基础上,对韧皮部运输和防御功能特别是碳水化合物装载和卸出机理进行了大量研究,克隆出一些特异基因,初步阐明了韧皮部装载的分子机制,取得可喜进展。本文综述了一些相关研究结果,并提出几个有待解决的问题。     

Companion-Cell Specific Localization of Sucrose Synthase in Zones of Phloem Loading and Unloading

An immunohistochemical approach was used in maize (Zea mays) and citrus (Citrus paradisi) to address the previously noted association between sucrose synthase and vascular bundles and to determine the localization of the low but detectable levels of sucrose synthase that remain in leaves after the import-export transition. Sucrose synthase protein was immunolocalized at the light microscope level using paraffin sections reacted with rabbit sucrose synthase polyclonal antisera and gold-conjugated goat anti-rabbit immunoglobulin G. Immunolabel was specifically observed in phloem companion cells of minor and intermediate veins in mature leaves of both species. Similar localization was apparent in the midrib of mature citrus leaves, with additional labeling in selected files of phloem parenchyma cells. A clear companion-cell specificity was evident in the phloem unloading zone of citrus fruit, where high activity of sucrose synthase has been demonstrated in vascular bundles during periods of rapid import. Sucrose synthase protein was not associated with adjacent cells surrounding the vascular strands in this tissue. The companion-cell specificity of sucrose synthase in phloem of both importing and exporting structures of these diverse species implies that this may be a widespread association and underscores its potential importance to the physiology of vascular bundles.     

Monitoring Phloem Unloading and Post-Phloem Transport by Microperfusion of Attached Wheat Grains

Phloem unloading and post-phloem transport in developing wheat (Triticum aestivum L.) grains were investigated by perfusing the endosperm cavities of attached grains. Relative unloading ratio (RUR) and the rate of sucrose release into the endosperm cavity (SRR) were calculated, respectively, from 14C import and from sucrose washout from the cavity. RUR and SRR continued at or near in vivo rates over a wide range of cavity sap osmolality (90 to approximately 500 milliosmolal) and sucrose concentration (14-430 mM) and for long times (29 h). These are much greater ranges than have been observed for the endosperm cavity in vivo (230-300 milliosmolal, and 40-120 mM, respectively), indicating that neither the cavity sap osmolality nor sucrose concentration are controlling factors for the rate of assimilate import into the cavity. The maintenance of in vivo transport rates over a wide range of conditions strongly implicates the role of transport processes within the maternal tissues of the wheat grain, rather than activities of the embryo or endosperm, in determining the rate of assimilate import into the grain. RUR was decreased by high concentrations of sucrose and sorbitol, but not of mannitol. By plasmolyzing some chalazal cells, sorbitol appeared to block symplastic transport across the crease tissues, but neither sucrose nor mannitol caused plasmolysis in maternal tissues of attached grains. The inhibition of RUR by KCN and carbonyl cyanide m-chlorophenyl (CCCP) and the continued import of sucrose into grains against its concentration gradient suggest that solute movement into the endosperm cavity might occur by active membrane transport. However, the evidence is weak, since KCN and CCCP appeared to act primarily on some aspect of symplastic (i.e. nonmembrane) transport. Also, sucrose could move from the endosperm cavity into the maternal tissues (i.e. opposite to the normal direction of sucrose movement), suggesting that transmembrane movement in the nucellus may be a reversible process. Pressure-driven flow into the grain could account for movement against a concentration gradient.     

菜豆茎韧皮部卸出的细胞途径以及激素的促进作用(英文)

通过缩小叶面积和去茎尖改变源库比率,以调节韧皮部卸出的途径,证明了韧皮部卸出的共质体与质外体途径的季节变化,和由对氯高汞苯磺酸所诱发的从质外体向共质体途径的转变,是与光合产物的输入有关。缩小叶面积而降低源库比率,能增加夏季生长植株茎韧皮部的质外体卸出,但对冬季生长植株无影响。去尖而增加源库比率,则促进共质体卸出。赤霉酸和激动素能促进共质体的横向转运,但对质外体转运无作用。当质外体为主要运输途径时,赤霉酸和激动素开启共质体途径。赤霉酸和激动素刺激光合产物,通过共质体从筛管一伴胞复合体向韧皮部薄壁纽胞输送,并可能在韧皮部薄壁细胞被动扩散到自由空间。由此可进一步说明蔗糖在激素处理部位自由空间的增加。     

Sink Metabolism in Tomato Fruit : II. Phloem Unloading and Sugar Uptake

Analysis of [³H]-(fructosyl)-sucrose translocation in tomato (Lycopersicon esculentum Mill.) indicates that phloem unloading in the fruit occurs, at least in part, to the apoplast followed by extracellular hydrolysis. Apoplastic sucrose, glucose, and fructose concentrations were estimated as 1 to 7, 12 to 49, and 8 to 63 millimolar, respectively in the tomato fruit pericarp tissue. Hexose concentrations were at least four-fold greater than sucrose at all developmental stages. Short-term uptake of [¹⁴C]sucrose, -glucose, and -fructose in tomato pericarp disks showed first order kinetics over the physiologically relevant concentration range. The uptake rate of [¹⁴C]-(glucosyl)-1′-fluorosucrose was identical to the rate of [¹⁴C]sucrose uptake, suggesting sucrose may be taken up directly without prior extracellular hydrolysis. Short-term uptake of all three sugars was insensitive to 10 micromolar carbonyl cyanide m-chlorophenylhydrazone and to 10 micromolar p-chloromercuribenzene sulfonic acid. However, long-term accumulation of glucose was sensitive to carbonyl cyanide m-chlorophenylhydrazone. Together these results suggest that although sucrose is at least partially hydrolyzed in the apoplast, sucrose may enter the metabolic carbohydrate pool directly. In addition, sugar uptake across the plasma membrane does not appear to be energy dependent, suggesting that sugar accumulation in the tomato fruit is driven by subsequent intracellular metabolism and/or active uptake at the tonoplast.     

Phloem loading in the tulip tree. Mechanisms and evolutionary implications

Minor vein ultrastructure and phloem loading were studied in leaves of the tulip tree (Liriodendron tulipifera; Magnoliaceae). Plasmodesmatal frequencies leading into minor vein companion cells are higher than in species known to load via the apoplast. However, these companion cells are not specialized as "intermediary cells" as they are in species in which the best evidence for symplastic phloem loading has been documented. Mesophyll cells plasmolyzed in 600 mM sorbitol, whereas sieve elements and companion cells did not plasmolyze even in 1.2 M sorbitol, indicating that solute accumulates in the phloem against a steep concentration gradient. Both [(14)C]sucrose and (14)C-labeled photo-assimilate accumulated in the minor vein network, as demonstrated by autoradiography. [(14)C]sucrose accumulation was prevented by p-chloromercuribenzenesulfonic acid, an inhibitor of sucrose-proton cotransport from the apoplast. p-Chloromercuribenzenesulfonic acid largely, but not entirely, inhibited exudation of radiolabeled photoassimilate. The evidence is most consistent with the presence of an apoplastic component to phloem loading in this species, contrary to speculation that the more basal members of the angiosperms load by an entirely symplastic mechanism.     

Water Flow Across the Sieve-Tube Boundary: Estimating Turgor and Some Implications for Phloem Loading and Unloading. II. Phloem in the Stem

Confirming a previous analysis by Lang (1974), it is concludedthat in tree trunks, phloem turgor and turgor gradients maybe estimated from osmotic pressure and osmotic-pressure gradients,respectively. The present analysis is an improvement becauseit is based on observed osmotic-pressure gradients rather thansupposed turgor gradients, and allowance is made for sucroseunloading and gradients of external water potential. It is concludedthat the rate of sucrose unloading in tree trunks must be lessthan 50 nmol m^–2 S m^–1. In small plants, higherrates of unloading (100 nmol m m^–2 S m^–1) and steeperconcentration gradients will lead to larger errors, but turgorpressures can still be estimated with acceptable accuracy. Oneshould be more cautious when considering turgor gradients insmall plants, although it seems likely that reasonable estimateswill still be obtained. Assuming plasmodesmatal transport throughan unconstricted cytoplasmic annulus, it is concluded that thesieve elements and their associated cells will sustain verysimilar turgor and osmotic pressures. Convection and diffusioncan both contribute significantly to plasmodesmatal sucroseunloading. Similarly, the plasmodesmatal volume flux will reflecta combination of pressure flow and osmosis. Water fluxes acrossthe sieve element plasmalemma and through the plasmodesmatacan be in opposite directions. It may be possible to assessthe extent of hydraulic coupling between the sieve elementsand their associated cells from studies of phloem water relations Phloem, turgor, osmotic pressure, plasmodesmata, phloem unloading, Munch hypothesis     

Water Flow Across the Sieve-Tube Boundary: Estimating Turgor and Some Implications for Phloem Loading and Unloading. III. Phloem in the Leaf

The theory described in Part I of this series is applied hereto the loading of water and sucrose into the sieve-element-companion-cell(se-cc) complexes of minor veins. It is concluded that symplasticphloem loading cannot account for large increases in osmoticpressure from mesophyll to se-cc complex or the dilution ofsieve tube sap which is evident in leaves. By contrast, loadingfrom the free space into a symplastically isolated se-cc complexcan account for both these observations. Within the se-cc complex,sucrose and water will be transported from the companion cellsto the sieve elements by a pressure-driven flow of solutionthrough the cytoplasmic annuli of plasmodesmata. The associatedchanges in turgor and osmotic pressure are small, and so these-cc complex can be regarded as a single compartment with respectto water potential. The assumption that this compartment isat water flux equilibrium will lead to significant overestimatesof turgor gradients. However, if the trans-membrane water potentialdifference and the external water potential are taken into account,the correlation of such gradients with sieve-element dimensionsand / or transport velocities provides one means of testingthe Munch hypothesis of phloem transport Phloem, turgor, osmotic pressure, plasmodesmata, Münch hypothesis, Phloem loading     

Phloem Unloading of Amino Acids at the Site of Attachment of Cuscuta europaea

By washing out ¹⁴C-solutes or ³H-solutes in 0.5 mm CaSO₄ during a period of 5 to 6 hours, the release of amino acids by excised stem segments of broad bean (Vicia faba L. cv Witkiem) was studied. Three hours after pulse labeling with l-valine, l-asparagine, or α-aminoisobutyric acid (AIB), hollow stem segments were excised from the plant and incubated in a washout solution.     

Calculation of Translocation Coefficients from Phloem Anatomy for use in Crop Models

Translocation coefficients, computed for unit ground area, arerequired in crop models as part of the simulation of the partitioningof assimilates. An equation for translocation is derived byconsidering pressure-driven flow and the physical dimensionsof the pathway in the phloem. Numerical estimates of the translocationcoefficients for grasses and trees are calculated using anatomicaldata. The values found are compatible with published rates oftranslocation in the phloem.Copyright 1995, 1999 Academic Press Model, Münch, phloem, sieve tube, translocation     

Developmental Changes in the Anatomy of the Sugarcane Stem in Relation to Phloem Unloading and Sucrose Storage

Transverse sections of immature and mature sugarcane internodes were investigated anatomically with white and fluorescence light microscopy. The pattern of lignification and suberization was tested histo-chemically. Lignification began in the xylem of vascular bundles and progressed through the sclerenchymatic bundle sheath into the storage parenchyma. Suberization began in parenchyma cells adjacent to vascular bundle sheaths and spread to the storage parenchyma and outer sheath cells. In mature internodes most of the storage parenchyma was lignified and suberized to a significant degree, except in portions of walls of isolated cells. The pattern of increasing lignification and suberization in maturing internodes more or less paralleled an increase of sucrose in stem tissue. In mature internodes having a high sucrose concentration, the vascular tissue was surrounded by thick-walled, lignified and suberized sclerenchyma cells. The apoplastic tracer dyes triso-dium 3-hydroxy-5,8,10-pyrenetrisulfonate (PTS) and amido black 10 B, fed into cut ends of the stalk, wereconfined to the vascular bundles in all internodes above the one that was cut — with no dye apparently in storage parenchyma tissue. Thus both structural and experimental evidence is consistent with vascular tissue being increasingly isolated from the storage parenchyma as maturation of the tissue proceeds. We conclude that in mature internodes the pathway for sugars from the phloem to the storage parenchyma is symplastic. The data suggest that an increasingly greater role for a symplastic pathway of sugar transfer occurs as the tissue undergoes lignification/suberization.     

Phloem Unloading within the Seed coat of Pisum sativum Observed using Surgically Modified Seeds

Phloem unloading in pea seed coats was observed by removingthe embryos from developing seeds and washing the attached coatswith a weakly buffered solution. The quantity of labelled photosynthateappearing in the washing solution varied immediately when thesolute concentration was changed, and is shown to be an osmoticresponse. This response is predicted by the Münch theoryof phloem transport with concentration dependent unloading.Respiratory inhibitors and the sulphydryl modifying reagentPCMBS had a slow effect upon the washout of tracer, which arrivedwithin the seed coat prior to inhibitor application, but completelystopped any washout of tracer arriving after its application.This time-course suggests that the inhibitors were not directlyinhibiting unloading, but preventing further tracer from enteringthe region of unloading within the seed coat. Phloem unloadingwithin the seed coats of Pisum appears to be passive and notdependent upon a PCMBS-sensitive carrier. Key words: Pisum sativum, seeds, phloem unloading     

Regulatory Mechanisms of Textural Changes in Ripening Fruits

Texture changes in ripening fruits influence consumer preference, fruit storability, transportability, shelf-life, and response to pathogen attack. Genetic regulatory factors as well as environmental conditions simultaneously affect texture changes in ripening fruit. Recent physiological and molecular studies provide insights into our knowledge and understanding of events and/or factors that contribute to changes in fruit texture, including softening and lignification. The roles of enzymes involved in modification and/or regulation of cell wall components as well as ethylene signaling components that play key roles in fruit textural changes during fruit ripening and storage will be presented and discussed. In addition, physical as well as chemical regulation of textural changes in ripening fruit will be explored.