52Fe Translocation in Barley as Monitored by a Positron-Emitting Tracer Imaging System (PETIS): Evidence for the Direct Translocation of Fe from Roots to Young Leaves via Phloem

The real-time translocation of iron (Fe) in barley (Hordeumvulgare L. cv. Ehimehadaka no. 1) was visualized using the positron-emittingtracer ⁵²Fe and a positron-emitting tracer imaging system (PETIS).PETIS allowed us to monitor Fe translocation in barley non-destructivelyunder various conditions. In all cases, ⁵²Fe first accumulatedat the basal part of the shoot, suggesting that this regionmay play an important role in Fe distribution in graminaceousplants. Fe-deficient barley showed greater translocation of⁵²Fe from roots to shoots than did Fe-sufficient barley, demonstratingthat Fe deficiency causes enhanced ⁵²Fe uptake and translocationto shoots. In the dark, translocation of ⁵²Fe to the youngestleaf was equivalent to or higher than that under the light condition,while the translocation of ⁵²Fe to the older leaves was decreased,in both Fe-deficient and Fe-sufficient barley. This suggeststhe possibility that the mechanism and/or pathway of Fe translocationto the youngest leaf may be different from that to the olderleaves. When phloem transport in the leaf was blocked by steamtreatment, ⁵²Fe translocation from the roots to older leaveswas not affected, while ⁵²Fe translocation to the youngest leafwas reduced, indicating that Fe is translocated to the youngestleaf via phloem in addition to xylem. We propose a novel modelin which root-absorbed Fe is translocated from the basal partof the shoots and/or roots to the youngest leaf via phloem ingraminaceous plants.     

Fe-deficiency stress response in Fe-deficiency resistant and susceptible subterranean clover: importance of induced H+ release

Plants can exhibit Fe-deficiency stress response when they areexposed to Fe-deficiency conditions. The relative importanceof the individual Fe-deficiency stress-response reactions, forexample, increased release of H⁺ from roots, enhanced root plasmamembrane-bound Fe³⁺ -reductase activity, and release of reductant,in Fe-deficiency resistance is not understood. To address thisproblem, the Fe-deficiency stress response of two cultivarsof subterranean clover (subclover), Koala (Trifolium brachycalycinumKatzn. and Morley) (Fe-deficiency resistant) and Karridale (T.subterraneum L.) (Fe-deficiency susceptible), were evaluated.The plants were cultured hydroponically at 0 (–Fe) and30 (+Fe) µM Fe³⁺ EDTA conditions. After 6 d Fe treatment,the –Fe Koala and Karridale decreased the pH of the nutrientsolution by 1.83 and 0.79 units, respectively, while the +Feplants increased the pH of the nutrient solution. The H⁺ -releaserate of the –Fe Koala determined 7 d after Fe treatmentinitiation was more than three times higher than that of the–Fe Karridale. The –Fe plants had a significantlyenhanced Fe³⁺ -reduction rate compared with the +Fe plants foreach cultivar, but the resistant cultivar did not exhibit ahigher root Fe³⁺ -reduction rate than the susceptible cultivarat each Fe treatment. Reductant release from the roots of subcloverwas negligible. These results indicate that Fe-deficiency-inducedH⁺ release may be the predominant factor influencing Fe-deficiencyresistance in subclover. Key words: Fe-deficiency, Fe³⁺ reduction, H⁺ release, stress response, Trifolium     

Transferrin stimulates iron absorption, exocytosis, and secretion in cultured intestinal cells

The cellularmechanism by which basolateral transferrin (Tf) produces an increase inapical-to-basolateral Fe flux in Caco-2 cells was analyzed. After apulse of ⁵⁹Fe from the apicalmedium, three types of basolateral⁵⁹Fe efflux were found: a⁵⁹Fe efflux that was independentof the presence of Tf in the basolateral medium, a⁵⁹Fe efflux in which⁵⁹Fe left the cell bound to Tf,and a Tf-dependent ⁵⁹Fe efflux inwhich ⁵⁹Fe came off the cell notbound to Tf. Furthermore, addition of Tf to the basolateral mediumdoubled the exocytosis rate of Tf and increased the secretion ofapolipoprotein A, a basolateral secretion marker. Both apotransferrinand Fe-containing Tf produced similar increases in⁵⁹Fe efflux, Tf exocytosis, andapolipoprotein A secretion. The Ca²⁺ channel inhibitor SKF-96365inhibited both the Tf-mediated increase in transepithelial Fe transportand the secretion of apolipoprotein A. Thus the activation oftransepithelial Fe transport by Tf seems to be mediated byCa²⁺ entry into the cells.     

The Behaviour of Iron Chelating Agents with Plants

The absorption by normal and iron-deficient bean plants of Feand of ethylene-diamine-bis-(orthohydroxyphenylacetic acid)(EDHPA) is described; and the metabolism in plants of this chelatingagent and of ethylenediaminetetra-acetic acid (EDTA) is examinedusing ¹⁴C-labelled compounds. Iron-deficient plants absorb Ferapidly and preferentially, absorption of the chelate itselfis slower but continuous. Normal plants absorb Fe and chelatein equimolar amounts. Bicarbonate ions in the culture solutioninhibit the preferential absorption of Fe by iron-deficientplants, equimolar amounts of Fe and chelate being taken up.These results are discussed in relation to apparently conflictingreports on Fe and chelate uptake in the literature. The recovery of ¹⁴C from plants treated with ¹⁴C-labelled chelatesdoes not reflect the true extent of the breakdown that has takenplace: with both EDTA and EDHPA less than half the ¹⁴C recoveredwas as unchanged chelate.     

Orientation of Pea Stem Mitochondrial H+-Pyrophosphatase and Its Different Characteristics from the Tonoplast Counterpart

Proton pumping pyrophosphatase (H⁺-PP_iase) of pea stem mitochondriaappears to be localized on the inner surface of the inner membrane.Aminohexanediphosphonate and dichloromethylenediphosphonateexert different inhibitory effects on this activity and on thatof tonoplast. Antibody raised against membrane-bound mitochondrialH⁺-PP_iase does not react with tonoplast vesicles. Thus, plantmitochondrial H⁺PP_iase seems to have a molecular structure differentfrom that of vacuolar H⁺-PP_iase. (Received August 2, 1996; Accepted October 18, 1996)     

Exposure of Dunaliella salina to Low Temperature Mimics the High Light-Induced Accumulation of Carotenoids and the Carotenoid Binding Protein (Cbr)

We report that growth of Dunaliella salina at either 13°C/150µmol m^–2s^–1 or 30°C/2,500 µmol m^–2s^–1 results in the accumulation of comparable levels ofcarotenoids and the zeaxanthin-binding protein, Cbr. We concludethat carotenoid and Cbr abundance in this green alga respondto changes in PSII ‘excitation pressure’ ratherthan to high light per se. (Received September 19, 1996; Accepted November 20, 1996)     

Determination of Levels of NO-3, NO-2 and NH+4 Ions in Leaves of Various Plants by Capillary Electrophoresis

Levels of NO^-₃, NO^-₂ and NH⁺₄ ions in leaves of six plant specieswere determined by capillary electrophoresis. Levels of NO^-₃ions differed by a factor of more than 350 in the six species.NO^-₂ and NH⁺₄ ions were detected in all species examined butat lower and more similar respective levels than NO^-₃ ions. (Received March 8, 1996; Accepted June 24, 1996)     

Uptake of zinc from chelate-buffered nutrient solutions by wheat genotypes differing in zinc efficiency

Zinc-efficient Triticum aestivum (cv. Warigal) and Zn-inefficientTriticum turgidum conv. durum (cv. Durati) were grown in chelate-buffered,complete nutrient solutions providing either deficient or sufficientZn supply. When transferred to fresh chelatebuffered nutrientsolutions containing a wide range of Zn supplies (0–1.28µmol m^–3 Zn²⁺ activity) for 24–48 h, bothgenotypes increased net Zn uptake linearly with an increasein solution Zn²⁺ activities. Zincefficient Warigal accumulatedZn at a greater rate than Zn-inefficient Durati. The greaterrate of net Zn uptake was observed by plants of both genotypeswhen pretreated at deficient Zn supply. Net loss of Zn to thesolution was higher in plants pretreated with sufficient Znand was inversely related to Zn²⁺ activity in the external solution.When continuously supplied with 40 nmol m^–3 Zn²⁺, netZn uptake by Zn-efficient Warigal was significantly greaterthan that of Zn-inefficient Durati, but the difference diminishedwith plant age. Shoot concentrations of Fe, Mn and Cu were higherwhen plants were grown at deficient than at sufficient Zn supply.The Zn-efficient genotype transported less Zn and Fe to shootsand had higher Fe concentrations in roots than the Zn-inefficientgenotype, supporting the hypothesis that Zn efficiency may beconnected with inefficient transport of Fe from roots to shootsand thus initiation of the Fe-deficiency response resultingin increased release of Zn- and Fe-binding phytosiderophores.It is concluded that differential Zn efficiency of wheat genotypesis at least partly due to a greater ability of efficient genotypesto accumulate Zn. Key words: Chelate-buffering, genotypes, micronutrients, Triticum spp., uptake, zinc efficiency     

Uptake of Neutral Red by the Vacuoles of a Green Alga, Micrasterias pinnatifida

Neutral red (NR) in the culture medium entered the vacuolesof a green alga, Micrasterias pinnatifida, at a higher rateat pH 8 than at pH 5. NR remained soluble in vacuoles of cellscultured at pH 5, while it precipitated and formed granulesin cells cultured at pH 8. The vacuoles of cells cultured atpH 8 contained fibrils, but those of cells cultured at pH 5did not. The amount of NR that entered the cells was markedlyreduced by the addition to the medium of nigericin at 10^-5M,monensin at 10^-5M, bafilo-mycin A₁ at 10^-5M, or ammonium chlorideat 50 mM. The formation of NR granules in vacuoles were stronglyinhibited and the disorganization of NR granules were acceleratedby the addition of nigericin at 10^-5M, or bafilomycin A₁ at10^-5M to the culture medium. The possibility is discussed thatNR which enters vacuoles might become positively charged (NRH⁺)by protons brought into vacuoles by proton pumps and that NRH⁺might combine with some negatively charged macromolecules toform aggregates or granules. (Received April 18, 1996; Accepted May 27, 1996)     

Studies on Mechanoperception in Characean Cells: Pharmacological Analysis

The mechanisms for generating receptor potentials and actionpotentials upon mechanical stimulation were studied in internodalcells of Chara. Receptor potentials and the subsequent actionpotentials could be generated even when the electrogenic protonpump was inhibited, indicating that the proton pump does notplay a central role in generating receptor potentials and actionpotentials. The involvement of Ca²⁺ and/or Cl^– channelsin both receptor and action potentials was suggested, basedon the equilibrium potentials of these ions across the plasmamembrane. Inhibitors of the Ca²⁺ channel, Cl^– channeland stretch-activated channel could not inhibit generation ofthe receptor potential. These findings suggested that the channelsinvolved in generating the receptor potential are insensitiveto these channel inhibitors, although all inhibitors significantlyinhibited the action potential. (Received July 26, 1996; Accepted November 19, 1996)     

Identification and Amino Acid Sequences of Tryptic Peptides of a Novel Ferredoxin-NADP+ Oxidoreductase from Rice

A 37-kDa protein purified from rice thylakoid membranes hasbeen identified as a ferredoxin-NADP⁺ oxidoreductase based onits catalysis of the reduction of nitro blue tetrazolium viaNADPH and its recognition by antibodies against ferredoxin-NADP⁺oxidoreductase. Amino acid sequences determined from trypticfragments of the enzyme further confirm the identity of theprotein and show the presence of unique sequences at the amino-terminus. (Received May 21, 1996; Accepted September 3, 1996)     

Rapid Reconstitution of Tonoplast H+-Translocating Pyrophosphatase from Cultured Rice Cells into Liposomes

We report the rapid and functional reconstitution of H⁺-pyrophosphatase(H⁺-PPase) from the tonoplast of cultured rice (Oryza sativaL.) cells to proteoliposomes. The CHAPS-solubilized H⁺-PPasewas incorporated into liposomes by gel-filtration. Both theactivities of PP_i-hy-drolysis and H⁺-pumping were influencedby the lipid-pro-tein ratio and cholesterol. (Received June 10, 1996; Accepted January 9, 1997)     

Suppression of Calcium Transport to Shoots by Root Restriction in Tomato Plants

Restriction of tomato roots by growth in small containers stronglysuppressed transport of ⁴⁵Ca²⁺ ions to new leaves and apices.Water transport, expressed on a leaf area basis, was marginallyreduced by root restriction, an indication that calcium transportwas more severely limited than water transport. (Received October 11, 1996; Accepted January 27, 1997)     

Dibromothymoquinone (DBMIB) Replaces the Function of QA at 77 K in the Isolated Photosystem II Reaction Center (D1-D2-Cytochrome b559) Complex: Difference Spectrum of the P680+(DBMIB-) State

Transient absorbance changes of the primary electron donor chlorophylla (P680) and acceptor pheophytin a (H) were measured at 77 Kby nanosecond laser spectroscopy in the D1-D2-cytochrome b₅₅₉photosystem II reaction center complex containing dibromomethylisopropylbenzoquinone (DBMIB). After the laser excitation of the reactioncenter in the presence of DBMIB, only the P680⁺-(DBMIB^-) statewas detected. P680⁺ mainly decayed with a t_1/e of 11 ms. Inthe absence of DBMIB, the excitation produced the P680⁺H^- radicalpair. The radical pair produced the triplet state (P680^T) witha t_1/e of 50 ns, and P680^T then decayed with a t_1/e of 2.1 ms.It was concluded that H_- was oxidized by DBMIB in a time rangefaster than the detecting time resolution (3.5 ns) even at 77K. The rapid oxidation of H^- by DBMIB was also confirmed bythe suppression of delayed fluorescence with a decay t_1/e of50 ns. The P680⁺(DBMIB^-)/P680(DBMIB) difference spectrum exhibiteda Q_y, band with a peak at 682 nm with a shoulder at 673 nm.The spectral shape was almost temperature insensitive between77 and 265 K. The feature of this spectrum in the wavelengthrange between 330 and 720 nm was compared with that of P680_T/P680or H^-/H at 77 K. (Received May 8, 1996; Accepted June 24, 1996)     

大豆初生根凯氏带对铜离子的通透性

采用在根内生成有色铜沉淀的方法研究大豆(Glycine max)初生根凯氏带对铜离子的通透性。用真空泵抽取浓度为200 μmol·L^–1 的CuSO₄溶液进入根中, 然后在重力作用下从根基部灌注400 μmol·L^–1的K₄[Fe(CN)₆]溶液, 两种物质在根内相遇即可产生棕色的Cu₂[Fe(CN)₆]沉淀, 根据沉淀的位置来确定铜离子所经过的途径。结果表明: Cu²⁺可以穿过内皮层凯氏带, 在木质部导管壁以及凯氏带至木质部之间的细胞壁处产生棕色沉淀, 侧根发生的部位也产生了大量的沉淀; 当抽取K₄[Fe(CN)₆]溶液后再灌注CuSO₄溶液, 发现Cu²⁺仍然可以穿过凯氏带, 并在凯氏带外侧以及外皮层细胞的细胞壁处产生棕色沉淀。研究结果证明凯氏带并不是一个可以完全阻止离子进出的完美屏障。     

Inhibition of Hill Reaction and Photophosphorylation by l,l,l-Trichloro-2,2-bis(p-chlorophenyl)ethane

The effect of DDT on DCIP and Fe(CN)₆^3– photoreductions,and cyclic and non-cyclic photo-phosphorylations, in some 30varieties of barley from widely different parts of the worldis reported. Whereas resistant barleys were not affected byDDT treatment, chloroplasts from treated susceptible barleysshowed parallel inhibitions of all the investigated aspectsof photosynthesis. However, in a few susceptible varieties inhibitionsof Fe(CN)₆^3– photoreduction or non-cyclic photophosphorylationwere not so pronounced. Possible reasons for these anomaliesare discussed; in particular earlier reports that DDT had noeffect on these latter photosynthetic activities may have beendue to the use of hypotonic media during chloroplast isolation.     

Distribution of Calanus species in Kongsfjorden, a glacial fjord in Svalbard

The distribution of Calanus species was investigated in Kongsfjordenin summer of 1996 and 1997. In both years Calanus finmarchicusand Calanus glacialis dominated, although the boreal C. finmarchicuswas more abundant than the Arctic C. glacialis in 1997. Thiscoincided with a 2°C higher water temperature at 50 m in1997, indicating stronger influence of Atlantic origin waterthat year. Advected Calanus finmarchicus occurred in deep andsubsurface layers of the outer fjord in 1996 (200 ind. m^-3,mainly CIII). A less abundant local population aggregated insurface layers of the inner fjord (100 ind. m^-3). Similarly,advected C. finmarchicus occurred in subsurface layers in 1997(446 ind. m^-3, mainly CIII and CIV) and a local population insurface layers (183 ind. m^-3, mainly CI). Calanus glacialisin 1996 aggregated as CII and CIII in the deep layers of theouter fjord (272 ind. m^-3), whereas CIII–CV were abundant(216 ind. m^-3) in cold surface waters of the inner fjord. In1997 C. glacialis (mostly CIII–CV) was more abundant inthe outer than in the inner part of the fjord (40 and 192 ind.m^-3, respectively). Within Kongsfjorden, Calanus finmarchicusneeds one year to complete its life cycle, whereas Calanus glacialisneeds two. Calanus hyperboreus seems to be an expatriate inthe fjord system.     

A Sodium Pump in the Plasma Membrane of the Marine Alga Heterosigma akashiwo

ATP-dependent transport of ²²Na⁺ into liposomes reconstitutedfrom plasma membrane proteins of Heterosigma akashiwo was examined.The apparent K^m values for transport of Na⁺ were 400 µMfor ATP and 7 mM for Na⁺. ATP-dependent transport of ²²Na⁺ wasnot inhibited by a protonophore or a membrane-permeable cationbut was inhibited by an inhibitor of P-type ATPases. (Received October 2, 1995; Accepted February 1, 1996)     

Influence of Complexation on the Uptake by Plants of Iron, Manganese, Copper and Zinc: I. EFFECT OF EDTA IN A MULTI-METAL AND COMPUTER SIMULATION STUDY

After growing barley (Hordeum vulgare L.) in nutrient solutionscontaining EDTA, uptake of the nutrient metals was determinedat three harvests and concentrations of the various chemicalspecies of each metal in the growth solutions was modelled bycomputer simulation. Complexation with EDTA had different effectson the uptake of the ions Fe³⁺, Mn²⁺, Cu²⁺, and Zn²⁺. At thehighest EDTA level (EDTA/Fe=2/l) the plants were chlorotic andgrowth was inhibited. This is attributed to a deficiency inZn rather than in Fe. The critical level of free Zn²⁺ requiredin nutrient solutions for healthy growth was found to be approximately10^–1010^–10 mol dm^–3, which is consistent withthat found by earlier workers for other plant species. Barleytolerated much lower levels of the free ions of copper and ironwithout exhibiting any obvious adverse effects. Key words: EDTA, micronutrients, trace metals, computer simulation, deficiencies, absorption, iron, manganese, copper, zinc     

The Level of mRNA Transcribed from psaL, Which Encodes a Subunit of Photosystem I, Is Increased by Cytokinin in Darkness in Etiolated Cotyledons of Cucumber

A cDNA clone for an mRNA whose level increased within 2 h ofthe start of treatment with N⁶-benzyladenine in etiolated cotyledonsof cucumber was isolated by differential hybridization. ThecDNA was homologous to psaL, which encodes subunit XI (PSI-L)of photosystem I. The accumulation of psaL mRNA was specificallyinduced by cytokinins or light. (Received April 10, 1996; Accepted July 31, 1996)