Potassium content and aperture in “intact” stomatal and epidermal cells ofCommelina communis L

Summary Measurements of potassium activity with a potassium-sensitive microelectrode have been made in the cells of the stomatal complex, and in epidermal cells, ofCommelina communis L., as a function of stomatal aperture. The estimated osmotic effects of the changing accumulation of potassium salts in the guard cell have been compared with the previous estimates of the osmotic changes required to open/close the pore. The results suggest that a significant fraction of the osmotic pressure of the guard cells, particularly when closed, is contributed by solutes other than potassium salts. The degree of potassium accumulation may determine the aperture of wide-open stomata, but the potassium changes in the early stages of opening are much too small to account for the osmotic changes required. The difference in potassium contents of intact and isolated guard cells is close to that required to overcome the previously estimated effect of subsidiary cell turgor on the water relations of the guard cell. In some tissue (but not in all) much more K is lost from epidermal cells than appears in other cells of the complex as the stomata open, and extracellular storage would be required.     

Fluxes of 86Rb+ in “isolated” guard cells of Vicia faba L.

Steady-state ⁸⁶Rb⁺ fluxes and contents were measured in stomatal guard cells of Vicia faba L., using isolated epidermal strips in which all cells other than the guard cells had been killed by a brief ultrasound treatment. Flux experiments were carried out for a range of stomatal apertures, achieved by incubation in light or dark, or on solutions containing different concentrations of RbCl (1–30 mM). At pH 5.5 the efflux curve of ⁸⁶Rb⁺ could be fitted to the sum of two exponential terms and it was possible to calculate compartmental contents and fluxes. The data indicate that steady-state stomatal opening is achieved by regulation of both influx and efflux. Salt accumulation was not always adequate to contribute the observed osmotic requirement for opening, and this may indicate that some other solute accounts for the discrepancy.Abbreviations and Symbols A amplitude - k rate constant - Mes 2-(N-morpholino)-ethanesulphonic acid - Q_T, Q_C, Q_V total, cytoplasmic, vacuolar chemical content - Q^* tracer content - Ø_P, Ø_l plasmalemma, tonoplast fluxes This work was supported by a Research Studentship from the Science and Engineering Research Council. I thank Professor E.A.C. MacRobbie for much helpful discussion and advice.     

Effects of light/dark and calcium-channel drugs on fluxes of 86Rb+ in “isolated” guard cells of Vicia faba L.

Experiments on ⁸⁶Rb⁺ fluxes were used to identify the changes in ionic state induced by transferring stomata from light to darkness. Guard-cell fluxes and contents were measured on isolated epidermal strips from Vicia faba L. in which all cells other than the guard cells had been killed by ultrasonic disruption. Closure of stomata in response to darkness was achieved by a large, transient stimulation of ⁸⁶Rb⁺ efflux from the guard cells, combined with a reduction of ion transfer from cytoplasm to vacuole, but there was little significant change in influx. Removal of blue but not red light appeared to trigger the flux responses associated with darkening. Attempts to inhibit the closing response (using methoxyverapamil, nifedipine and bepridil, compounds possessing activity against the calcium channels of animal cells) were mostly unsuccessful and the significance of this result is discussed.Abbreviations and Symbols A amplitude - K rate constant - Mes 2-(N-morpholino)ethanesulfonic acid - Q ^* tracer content - s specific activity - R rate of trace loss - Q _T, Q _C, Q _V total, cytoplasmic and vacuolar contents - flux This work was supported by a Research Studentship from the Science and Engineering Research Council. I thank Professor E.A.C. MacRobbie for much helpful discussion and advice.     

γ-Tubulin is associated with a cortical-microtubule-organizing zone in the developing guard cells of Allium cepa L.

A key event in the differentiation of elliptically shaped guard cells such as those in Allium is the formation of a radial array of cortical microtubules (Mts) which, by controlling the orientation of wall microfibrils, plays an important role in cell shaping. Previous experiments strongly indicated that the array is nucleated in a zone adjacent to the new ventral wall soon after cytokinesis. In order to further clarify the function of this zone, we performed dual immunolocalizations on Allium guard cells with anti--tubulin, to detect Mts, and an antibody to -tubulin, a protein known to be present at Mt-organizing centers in other species and recently identified in plants as well. -Tubulin antibody stained the cortical zone adjacent to the ventral wall, while little or no fluorescence was present elsewhere along the radial Mt array or at other sites in the cell. The antibody also stained the mitotic poles and phragmoplast in guard mother cells, as it does in other material. No staining was seen when the primary antibody was omitted. The results are consistent with nucleation of the radial array at a cortical-Mt-organizing zone next to the ventral wall, and set the stage for more in-depth studies on the spatial and temporal control of Mt formation in differentiating cells.Abbreviations CLSM confocal laser scanning microscope - FITC fluorescein isothiocyanate - Mt microtubule - MTOC microtubule-organizing center This work was supported by National Science Foundation grant DCB-9019285 to B.A.P., National Institutes of Health (NS30009) and American Cancer Society (CD6255) grants to H.C.J., and a University of Georgia Graduate School Assistantship to B.L. We thank Dr. Mark Farmer and the University of Georgia Center for Advanced Ultrastructural Research for the use of the confocal microscope.     

Chlorophyll “a” content in cells of Antarctic phytoplankton

Material was collected from the Weddell Sea and the Bransfield Strait in January/April 1989. Data on size-taxonomic composition and biomass of phytoplankton communities and Ch1 concentration were obtained to estimate the chlorophyll a (Ch1) cell content. Single cell fluorescence measured microscopically was used as a relative index of cellular Ch1 content of individual species. The relationship between the species composition of the algal communities and the ratio of phytoplankton carbonCh1 concentration (CCh1) was found. Due to changes in species composition the average CCh1 ratio in March/April (56) was half that in January/February (115). The CCh1 ratio ranged from 24 to 215 (mean=101) in the upper mixed layer and from 14 to 69 (mean=37) in the pycnocline region. The distribution of cellular Ch1 within individual species showed lower heterogeneity in the mixed layer in comparison with that in the pycnocline and below. Below the mixed layer, populations consisted partly of dead cells with very low pigment content, while other cells had greatly increased cellular Ch1. At several stations this cellular Ch1 increase led to the formation of a deep Ch1 maximum.     

Flavonoid content and antioxidant activity of Artemisia vulgaris L. “hairy” roots

We investigated the effect of Agrobacterium rhizogenes-mediated transformation on antioxidant activity of Artemisia vulgaris “hairy” roots. It appeared that transformation may increase flavonoid content as well as DPPH-scavenging activity and ability to reduce Fe³⁺ as compared to the non-transformed plants. Some “hairy” roots accumulated flavonoids up to 73.1?±?10.6?mg RE/g DW (while the amount of flavonoids in the leaves of non-transformed plants was up to 49.4?±?5.0?mg RE/g DW). DPPH-scavenging activity of some “hairy” root lines was 3–3.8 times higher than such one of the roots of the control plants. The Fe³⁺-reducing power of most transgenic root extracts exceeded such power of the extracts of the roots of the control plants. The decrease in SOD activity was found in the most “hairy” root lines compared to the control roots. The increase of flavonoid content correlated with the increase of ability of extracts to scavenge DPPH*- radical and Fe³⁺ - reducing power. No correlation between SOD activity of extracts and concentration of flavonoids was found (p?≥?0.2).Thus, transformation has led to the alteration in flavonoid accumulation and antioxidant activity in A. vulgaris “hairy” roots. Transgenic roots with high-antioxidant properties can be selected after A. rhizogenes-mediated transformation.     

Ion mapping in plant cells – methods and applications in signal transduction research

 This review covers both methodical aspects and actual applications of ion imaging techniques in plant cell signal research. The methodological section explains the basic principles of fluorescence ion imaging, the impact of modern developments in fluorescence microscopy and introduces the most important fluorescence probes including aequorin and other photoproteins. It critically comments on loading strategies, intracellular compartmentation of probes and calibration procedures. The second part compiles actual research areas where the application of ion imaging procedures has gained substantial achievements and helped to establish new concepts of calcium- and pH-dependent signalling. Examples comprise the hormonal control of stomatal movements, effects of gibberellic and abscisic acids in aleurone cells, elicitation of phytoalexin production, cytosolic pH and cell development, and signatures of Ca²⁺ as a universal signal in plant cells. Received: 26 May 1999 / Accepted: 2 August 1999     

Say “NO” to ABA signaling in guard cells by S-nitrosylation of OST1

<正>Nitric oxide(NO),a gaseous compound,plays important roles in plant immunity,abiotic stress response and plant development[1].In plants,NO is synthesized through either oxidative or reductive route that is dependent on the nitrate reductases(NADH)1(NIA1)and NIA2.NO bioactivity is realized through redox-based post-translational protein     

Differences in the floral anthocyanin content of violet–blue flowers of Vinca minor L. and V. major L. (Apocynaceae)

Two novel delphinidin 3-(tri or di)-glycoside-7-glycosides were isolated from the violet–blue flowers of Vinca minor L. and V. major L. (Family: Apocynaceae), and determined to be delphinidin 3-O-[2-O-(β-xylopyranosyl)-6-O-(α-rhamnopyranosyl)-β-galactopyranoside]-7-O-(α-rhamnopyranoside) [= delpphinidin 3-(2^G-xylosylrobinobioside)-7-rhamnoside] as major floral anthocyanin of V. minor and delphinidin 3-O-[6-O-(α-rhamnopyranosyl)-β-galactopyranoside]-7-O-(α-rhamnopyranoside) [= delpphinidin 3-robinobioside-7-rhamnoside] as major floral anthocyanin of V. major by chemical and spectroscopic methods. In addition, chlorogenic acid and kaempferol 3-O-[6-O-(α-rhamnopyranosyl)-β-galactopyranoside]-7-O-(α-rhamnopyranoside) [= kaempferol 3-robinobioside-7-rhamnoside (robinin)] were identified in these flowers. In this paper, the relation between the structure of floral anthocyanins and classification of Vinca species was discussed.     

Polysaccharide isolated from seeds of Plantago asiatica L. induces maturation of dendritic cells through MAPK and NF-κB pathway

Plantago species are used as traditional medicine in Asian and Europe. Polysaccharide isolated from the seeds of Plantago asiatica L. could stimulate maturation transformation of bone-marrow derived dendritic cells (DCs). We found that blocking p38, ERK1/2 and JNK MAPK signal transduction could significantly decreased the PLP-2 induced expression of MHC II, CD86 surface molecules on DCs. Blocking p38 and JNK signal also significantly inhibited the cytokine secretion of TNF-α and IL-12p70 as well, while blocking ERK1/2 signal only decreased the secretion of TNF-α. Meanwhile, DCs in the three MAPK signal-blocking groups showed dramatically attenuated effects on stimulating proliferation of T lymphocytes. Similarly, blocking signal transduction of NF-κB pathway also significantly impaired the phenotypic and functional maturation development of DCs induced by PLP-2. These data suggest that MAPK and NF-κB pathway mediates the PLP-induced maturation on DCs. Especially, among the three MAPK pathways, activation of JNK signal transduction is the most important for DCs development after PLP-2 incubation. And PLP-2 may activate the MAPK and NF-κB pathway by triggering toll-like receptor 4 on DCs.     

Shooting of sporidium by “gun” cells in Haptoglossa heterospora and H. zoospora and secondary zoospore formation in H. zoospora

Haptoglossa spp. (Lagenidiales, Oomycetes) have been known to parasitize microscopic animals by means of a “gun” cell that shoots an infection cell, named the sporidium, into the body of the animal. A thallus grown from the sporidium changes into a zoosporangium at maturation to produce a number of zoospores that encyst after a swarming period, and the resulting cysts germinate to produce gun cells. In Haptoglossa zoospora, endoparasitic in nematodes, the cysts of primary zoospores that swam for about 5min did not develop gun cells but produced secondary zoospores that swam for about 3h. After encystment of the secondary zoospores, each secondary cyst germinated to produce a gun cell. In the present study, the secondary zoospores of the genus Haptoglossa could be recorded with a videocassette recorder for the first time. The videocassette recording also revealed the infection of a nematodes by H. zoospora and H. heterospora to be composed of two steps of injection of a sporidium by the gun cell, in which the gun cell came in contact with the cuticle of a nematode and produced a spherical adhesorium on the tip of the cell in 0.07–0.1 s in both species. The adhesorium was ~2 μm in H. zoospora and ~4 μm in H. heterospora. When the adhesorium inflated to full size, it shot the sporidium into the nematode's body in 0.5–0.65 s and in 0.2–0.5 (or rarely 1.0) s in H. zoospora and H. heterospora, respectively. After shooting, the empty gun cell with an empty cyst case was separated from the cuticle immediately in both species.     

Glycogen synthetase in the sea mussel Mytilus edulis L.—II. Seasonal changes in glycogen content and glycogen synthetase activity in the mantle tissue

• 1.1. The glycogen content of the mantle tissue reached a maximum in the summer (May–July) with levels of 41.0–53.5% of the dry tissue weight.
• 2.2. Seasonal changes in glycogen synthetase activity showed that the I-activity (independent of G6P) increased up to 10-fold in June as compared with December. The measured I-activity of glycogen synthetase was sufficient to account for the accumulation of mantle glycogen in the summer.
• 3.3. The I-activity of glycogen synthetase declined rapidly in July of each year. A possible role for the inhibition of glycogen synthetase by high levels of tissue glycogen is suggested.
• 4.4. The I-activity in the mantle tissue of mussels on the shore was higher than that for animals starved in the laboratory for 2–3 days. The differences were minimal in early May but increased markedly in late May–July. Starved mussels returned to the shore showed an increase in I-activity of glycogen synthetase.
• 5.5. Injection of 30 μmol glucose into the adductor muscle increased the concentration of glucose in the mantle fluid to 2.0–2.5 mM. A similar injection of 60 μ mol glucose resulted in a time-dependent increase in the I-activity of glycogen synthetase.
• 6.6. Injection of mussels with mammalian insulin or anti-insulin serum had no effect on the activity of glycogen synthetase. Our results are at variance with those of other workers who have used the mammalian hormone in molluscan studies (see Discussion).

     

A cytochemical and ultrastructural study of the “S.I.F.” cells in cat sympathetic ganglia

Summary According to the hypothesis of Eccles and Libet, the small intensely fluorescent cells (S.I.F. cells) in the sympathetic ganglion would represent an essential element in the inhibition of the principal neuron. As a contribution to the study of this important problem, we have investigated serial sections in superior cervical (S.C.G.) and celiac (C.G.) ganglia of the cat, a species that has not been extensively studied up to now, both by fluorescence and electron microscopy. We have shown that the S.I.F. cells are three times fewer in the cat S.C.G. than in the rat S.C.G. There are five times more S.I.F. cells in the C.G. of the cat than in the S.C.G. of the same species. Moreover we have described two types of S.I.F. cells.Type I is composed of cells characterized by highly polymorphous large dense-cored vesicles. These cells lack processes and are grouped in clusters centered on fenestrated capillaries. They could be endocrine function cells. Type II is formed of isolated cells which exibit long processes and establish synaptic junctions with the dendrites of the principal neurons. In this case, the dense-cored vesicles are very regular and much smaller. These cells could be equivalent to interneurons. Type I very strongly predominates in the S.C.G. and C.G. of the cat where it represents more than 90% of the S.I.F. cell total observed by fluorescence microscopy. A priori such a quantitative and qualitative heterogeneity hardly consistent with Eccles and Libet's hypothesis based on the existence of dopaminergic interneurons only, allows the question to be raised as to the functional significance of the S.I.F. cells in ganglion physiology. The notion of modulation of ganglionic transmission does not seem to be quiered by these new data but could be founded on different forms of action embodied in the broader conception of the neuromodulation phenomenon.     

Behaviour in culture of isolated protoplasts from “Paul's scarlet” rose suspension culture cells

Summary Protoplasts were enzymatically isolated from Paul's scarlet rose suspension culture cells. They were cultured in medium similar to that used to culture the cells from which they were isolated with the addition of sucrose as an osmotic stabiliser. They were studied by light and electron microscopy and their changes in size and number per culture were recorded. Expansion was greater when the protoplasts were cultured in medium plus 12% sucrose than with 24% sucrose. Budding was observed. In medium plus 12% sucrose about 45% of the protoplasts divided but in medium plus 24% sucrose far fewer divided. Cytokinesis was abnormal: the phragmoplast disappeared soon after cytokinesis began and the cell plate became a groove and then a fibril-lined or filled tongue which progressed across the vacuole, unconnected by strands to other parts of the protoplast. The wall regenerated after several days culture in medium plus 12% sucrose fluoresced with calcofluor. The wall regenerated in medium with 24% sucrose fluoresced usually only after several weeks culture. Cytokinesis hastened formation of a wall fluorescing with calcofluor. In the electron microscope the wall was seen to contain fibrils and non-fibrillar material. The latter was the minor component in medium plus 12% sucrose but was usually the major component in medium plus 24% sucrose. The growth in plasmolysing and nonplasmolysing medium of the cells from which protoplasts are isolated was also studied.It appears that loss of the wall alters the potential of protoplasts to expand and possibly also to regenerate a wall and to divide. Wall regeneration is initially linked with expansion and cytokinesis. Osmotic pressure of the external medium is also an important factor.This work formed part of a thesis by one of us (R.S.P.) approved for the degree of Ph. D. in the University of Nottingham. The work was supported by the Agricultural Research Council.     

Stomatal movement in response to long distance-communicated signals initiated by heat shock in partial roots of Commelina communis L.

It is well known that animals are able to positively respond to environmental stimuli, thereby avoiding or reducing the possible injures or impacts on them, and the base for the rapid and positive responses is long- distance signal transmission mediated b…