Lemon maturation causes anatomical and biochemical changes at the flavedo tissue level

Plants mobilize the photosynthates by three transport pathways: apoplastic, symplastic through plasmodesmata (PD), and transcellular. In flavedo of postharvest mature lemons, a high activity of cell wall-bound invertase (WI), an enzyme associated with transcellular transport of monosaccharides, has been detected. In order to elucidate whether this high enzymatic activity is related to restricted transport in the symplastic pathway with fruit maturation, the aim of the present work was to compare anatomical and biochemical parameters in peel tissues of immature and mature lemons. Anatomical structure focusing on cell walls, callose deposition, WI activity, and sucrose content were analyzed in peel tissues of immature and mature lemons. The parenchyma of flavedo tissue of immature lemons presented an elevated number of primary pit fields (PPF). These PPF, associated to PD or cell wall interruptions, had the appearance of a string of beads. However, in mature lemons, the number of PPF was scarce due to callose deposition. WI activity and apoplastic sucrose content increased significantly in flavedo of mature lemons in comparison to immature lemons. Present findings lay structural and functional bases relevant to understand differences between immature and mature lemons, which would help to design agricultural practices in pre- and post-harvest management.

Symplasmic phloem unloading and post-phloem transport during bamboo internode elongation

In traditional opinions, no radial transportation was considered to occur in the bamboo internodes but was usually considered to occur in the nodes. Few studies have involved the phloem unloading and post-phloem transport pathways in the rapid elongating bamboo shoots. Our observations indicated a symplastic pathway in phloem unloading and post-unloading pathways in the culms of Fargesiayunnanensis Hsueh et Yi, based on a 5,6-carboxyfluorescein diacetate tracing experiment. Significant lignification and suberinization in fiber and parenchyma cell walls in maturing internodes blocked the apoplastic transport. Assimilates were transported out of the vascular bundles in four directions in the inner zones but in two directions in the outer zones via the continuum of parenchyma cells. In transverse sections, assimilates were outward transported from the inner zones to the outer zones. Assimilates transport velocities varied with time, with the highest values at 0):00 h, which were affected by water transport. The assimilate transport from the adult culms to the young shoots also varied with the developmental degree of bamboo shoots, with the highest transport velocities in the rapidly elongating internodes. The localization of sucrose, glucose, starch grains and the related enzymes reconfirmed that the parenchyma cells in and around the vascular bundles constituted a symplastic pathway for the radial transport of sugars and were the main sites for sugar metabolism. The parenchyma cells functioned as the ‘rays’ for the radial transport in and between vascular bundles in bamboo internodes. These results systematically revealed the transport mechanism of assimilate and water in the elongating bamboo shoots.

Pearl millet (Pennisetum glaucum) contrasting for the transpiration response to vapour pressure deficit also differ in their dependence on the symplastic and apoplastic water transport pathways

Genotypic differences in transpiration rate responses to high vapour pressure deficit (VPD) was earlier reported. Here we tested the hypothesis that this limitation could relate to different degrees of dependence on the apoplastic (spaces between cells), and symplastic water transport pathways (through cells via aquaporin-facilitated transport), which are known to have different hydraulic conductivities. The low transpiration rate (Tr) genotype PRLT 2/89/33 either restricted its transpiration under high VPD, or was more sensitive to VPD than H77/833-2, when grown hydroponically or in soil. The slope of the transpiration response to an ascending series of VPD was lower in whole plants than in de-rooted shoots. In addition, the transpiration response of detached leaves to moderately high VPD (2.67 kPa), normalised against leaves exposed to constant VPD (1.27 kPa), was similar in low and high Tr genotypes. This suggested that roots hydraulics were a substantial limitation to water flow in pearl millet, especially under high VPD. The dependence on the apoplastic and symplastic water transport pathways was investigated by assessing the transpiration response of plants treated with inhibitors specific to the AQP-mediated symplastic pathway (AgNO3 and H2O2) and to the apoplastic pathway (precipitates of Cu(Fe(CN)6) or Cu(CuFe(CN)6)). When CuSO4 alone was used, Cu ions caused an inhibition of transpiration in both genotypes and more so in H77/833-2. The transpiration of high Tr H77/833-2 was decreased more by AQP inhibitors under low VPD (1.8 kPa) than in PRLT 2/89/33, whereas under high VPD (4.2 kPa), the transpiration of PRLT 2/89/33 was decreased more by AQP inhibitors than in H77/833-2. The transpiration rate of detached leaves from H77/833-2 when treated with AgNO3 decreased more than in PRLT 2/89/33. Although the root hydraulic conductivity of both genotypes was similar, it decreased more upon the application of a symplastic inhibitor in H77/833-2. The transpiration of low Tr PRLT 2/89/33 was decreased more by apoplastic inhibitors under both low and high VPD. Then the hydraulic conductivity decreased more upon the application of an apoplastic inhibitor in PRLT 2/89/33. In conclusion, both pathways contributed to water transport, and their contribution varied with environmental conditions and genotypes. Roots were a main source of hydraulic limitation in these genotypes of pearl millet, although a leaf limitation was not excluded. The similarity between genotypes in root hydraulic conductivity under normal conditions also suggests changes in this conductivity upon changes in the evaporative demand. The low Tr genotype depended more on the apoplastic pathway for water transport, whereas the high Tr genotype depended on both pathway, may be by ‘tuning-up’ the symplastic pathway under high transpiration demand, very likely via the involvement of aquaporins.

Pathway and control of sucrose import into initiating cotton fibre cells

This paper originates from a presentation at the International Conference on Assimilate Transport and Partitioning, Newcastle, NSW, August 1999 Our aim is to unravel the mechanisms controlling fibre cell initiation from the epidermis of cotton (Gossypium hirsutum L.) ovules. We compared the development of fibres and trichomes in wild type cotton and a fibreless seed (fls) mutant, and determined the cellular pathway of sucrose transport into fibre initials on the day of anthesis. Although fibre initiation is inhibited in the fls mutant, leading to the fibreless phenotype, trichome development in other parts of the plant is normal. Confocal imaging analysis revealed that the fluorescent molecule, 5(6)-carboxyfluorescein, which is transported symplastically, moved readily from the integument phloem into initiating fibres. Plasmolysis studies showed that the fibre initials and adjacent non-initiating ovule epidermal cells have similar osmotic potential. Immunolocalisation analysis showed the absence of sucrose transporter proteins in the initiating fibre, but their abundance in the transfer cell precursors at the innermost integument. These results (i) demonstrate that fibre cell initiation is controlled by unique mechanism(s) that differ from that for normal trichome development; (ii) show a symplastic pathway of sucrose import into initiating fibres and strengthen the current opinion that sucrose synthase is likely to be the key enzyme mobilising sucrose into initiating fibres; and (iii) suggest that the initial protrusion of the fibre cells above the ovule surface is largely achieved by increased cell wall extensibility rather than higher turgor as is commonly thought.

Ion distribution in cereal leaves: pathways and mechanisms

Measurement of ion concentrations in the vacuoles of different cell types in cereal leaves using a variety of techniques indicates that ions are differentially distributed between different cell types. Thus mesophyll cells are enriched in P but contain relatively little Ca 2+ or Cl - , whereas the reverse is true for epidermal cells. Solutes reach the leaf via the transpiration stream and we consider three possible pathways which they could follow from the xylem to leaf cells. The first is a fully apoplastic mesophyll pathway in which both water and solutes move together through the leaf apoplast passing bundle sheath, mesophyll and epidermis in turn. The second is a partly symplastic mesophyll pathway in which ions and water pass into the symplast at the mestome/bundle sheath cells. Water continues to sites of evaporation via either a transcellular or symplastic pathway, but ions may be secreted back to the mesophyll apoplast and move to the epidermis along an extracellular route. The third is a vein extension pathway which provides a diffusional pathway for ions to the epidermis. A testable hypothesis for the roles of the pathways in supplying solutes to the mesophyll and epidermis is proposed and the implications of each of these pathways for transport systems in individual cell types is discussed.

The anatomy of the leaf of red pine, Pinus resinosa. II. Vascular tissues

The anatomy and ultrastructure of the endodermis and enclosed vascular tissues of the midregion of the mature secondary needle-leaf of Pinus resinosa are described. Within the uniseriate endodermis are two vascular traces surrounded by transfusion tissue. The endodermal cells have differentially thickened walls which lack Casparian strips but are lignified. Plasmodesmata traversing pit regions form a symplastic interconnection between mesophyll, endodermal, and transfusion parenchyma cells. In the lateral bundle region plasmodesmata extend this symplastic pathway across the cell walls of subjacent transfusion parenchyma and richly protoplasmic albuminous cells to the metaphloem. Four distinct types of transfusion tracheids have been defined on the basis of cell shape and location. Transfusion tracheids in the lateral bundle regions form direct radial connections between metaxylem and endodermis.

Leaf free space analysis and vein loading in Cucurbita pepo

To determine the role of the apoplast in the loading of the minor veins of Cucurbita pepo L., leaves were examined either for the ability to release selectively the transport sugars, sucrose and stachyose, from the metabolic space (MS) to the free space (FS) or to accumulate selectively exogenously fed transport sugars from the FS into the minor veins. FS extracts collected by vacuum infiltration and centrifugation of specially trimmed leaves were found to contain all sugars also present in ethanol extracts of the MS of the same leaves, but in amounts two orders of magnitude lower. Similarly, 14C activity in FS extracts from 14CO2-labelled leaves, which was distributed between all sugars, amino acids, and organic acids also found labelled in the MS, was nearly three orders of magnitude lower than in MS ethanol extracts. No excess of the transport sugars was evident in FS extracts. When 14C-labelled sucrose, stachyose, or galactose (all 1–2 mM) were infiltrated into the FS a rapid accumulation of these sugars into the mesophyll was apparent, as all sugars were rapidly interconverted into the various sugars normally found labelled in the MS after exposure of leaves to 14CO2. However, while leaves exposed to 14CO2 translocated label predominantly in the form of [14C]stachyose, label in exogenously fed leaves was translocated predominantly as [14C]sucrose, irrespective of the nature of the fed sugar. Exogenously fed transport sugars did not therefore appear to be taken up directly into the minor veins from the FS.The absence of significant levels of transport sugars in the FS as well as the failure of C. pepo leaves to load any appreciable amount of exogenously supplied [14C]stachyose, the predominant transport sugar in this species, would tend to preclude both a selective release of transport sugars into the apoplast and a selective uptake of transport sugars from the apoplast into the minor veins. A completely symplastic pathway for minor vein loading in C. pepo leaves therefore remains a possibility.

The structure of syncytia induced by the phytoparasitic nematode Nacobbus aberrans in tomato roots, and the possible role of plasmodesmata in their nutrition

The structure of syncytia induced within galls in tomato roots by the false root-knot nematode Nacobbus aberrans has been examined by light and electron microscopy. A syncytium develops by breakdown or individual cell walls, which allows movement of cytoplasmic contents between transformed cells. The wall breakdown takes place at pit fields, where the plasmodesmata may be protected from digestion until the surrounding wall is removed. Numerous sieve elements differentiate in the cells outside the syncytium. These sieve elements, and also plasmodesmata in pit fields, are demonstrated by fluorescence microscopy. The possibility of a symplastic pathway of solute movement from the phloem to the syncytium is suggested. A massive accumulation of starch occurs in the gall cells and syncytial cells, which may be related to the proliferation of phloem. Wall ingrowths typical of transfer cells are absent, and a comparative survey of the structure and mode of solute entry into nematode-transformed cells in which ingrowths are present or absent is presented.