bluuebunny/crossref_metadata_2025 huggingface dataset:
Multiple mechanisms control the expression of abscisic acid (ABA)‐requiring genes in tomato plants exposed to soil water deficit
Authors: Cohen A. , Moses M. S. , Plant Á. L. , Bray E. A.
Published: March 2003

Elevated levels of abscisic acid (ABA) are required for the expression of several genes during water deficit. The expression patterns of the ABA‐requiring genes, le4, le16, le25 and his1‐s, differed during a 4 d period of water deficit and a 24 h recovery period in intact tomato plants. During water deficit, a strong correlation between ABA content and mRNA accumulation was observed for le16, with only a moderate or poor correlation observed between the accumulation of ABA and the other mRNAs. The regulation of le4 and le16 can be accounted for by an increase in transcription rate, while components of post‐transcriptional regulation are implicated in the regulation of the le25 and his1‐s genes. A strong correlation between leaf ABA content and the transcription rate of le4 and le16 during the stress and recovery periods was observed. Protein synthesis was not necessary for the expression of the four genes. However, the mRNA content of le25 increased in the presence of translation inhibitors plus ABA, implicating inhibition of a transcriptional repressor. The regulation of gene expression in response to stress is complex; although ABA was required for the regulation of the genes in this study, there must be additional control mechanisms that interact with components of the ABA regulatory machinery.
Adaptive nature of the transition phases in development: the case of Sorghum bicolor
Authors: Amzallag G. N.
Published: March 2003

An increase in tolerance to salinity is induced in Sorghum bicolor by exposure to a sublethal concentration of NaCl during early vegetative development. The phase of competence for induction of this response, termed salt adaptation, is well defined in time and it coincides with the emergence of the first adventitious roots. The link between these events was investigated. Competence for salt adaptation varies among genotypes. It is shown that competence is especially high for genotypes in which the link between the seminal root and the shoot is reduced during emergence of the adventitious root. These data relate the capacity for salt adaptation with development in the absence of NaCl, suggesting that: (i) functional integration of the adventitious roots within the whole plant has an adaptive nature in normal development; (ii) salt adaptation results from an integration of the environmental constraint (NaCl) during this developmental readjustment. It is concluded that perturbations generated by emergence of a new organ are the cause of rapid variations in sensitivity required to open a competence window.
Phosphate uptake in Chara: membrane transport via Na/Pi cotransport
Authors: Reid R. J. , Mimura T. , Ohsumi Y. , Walker N. A. , Smith F. A.
Published: March 2003

Phosphate uptake in the freshwater charophyte plant Chara corallina was found to be strongly dependent on the presence of Na in the external medium. Based on the reciprocal stimulations of 32Pi uptake by Na and 22Na uptake by Pi, the logical mechanism for Pi uptake appears to be a nNa/Pi symport with a half‐maximal stimulation (Km) for Na of approximately 300 μM and a Km for Pi of approximately 10 μM. Comparison of the stimulations of 32Pi and 22Na influxes at pH 6 gives a stoichiometry of Na : Pi of 5·68. The reduction in Pi influx with increasing pH is consistent with the transported species being the monovalent H2PO4−. In voltage‐clamp experiments, currents elicited by Pi in the presence of Na were equivalent to an influx of positive charge which exceeded the measured influxes of 32P by a factor of 6·26. Intracellular perfusion was used to examine the dependence of Pi influx on ATP and Na. In perfused cells, Pi influx was low when ATP was absent from the internal medium or Na was absent from the external medium. Addition of ATP alone had little effect whereas addition of Na alone increased the 32Pi influx slightly. Addition of both ATP and Na together restored Pi influx to rates comparable to those of intact cells. It is suggested that the ATP is required for membrane hyperpolarization which in turn drives the highly electrogenic flux of Pi with up to 6 Na. However, consideration of the electrochemical potential differences for Na and Pi at pH less than 6 shows that nNa/Pi would not be feasible. It is suggested that at low pH, H+ may substitute for Na.
Susceptibility to photoinhibition of three deciduous broadleaf tree species with different successional traits raised under various light regimes
Authors: Kitao M. , Lei T. T. , Koike T. , Tobita H. , Maruyama Y.
Published: March 2003

The susceptibility to photoinhibition of tree species from three different successional stages were examined using chlorophyll fluorescence and gas exchange techniques. The three deciduous broadleaf tree species were Betula platyphylla var. japonica, pioneer and early successional, Quercus mongolica, intermediate shade‐tolerant and mid‐successional, and Acer mono, shade‐tolerant and late successional. Tree seedlings were raised under three light regimes: full sunlight (open), 10% full sun, and 5% full sun. Susceptibility to photoinhibition was assessed on the basis of the recovery kinetics of the ratio of vaviable to maximum fluorescence (Fv/Fm) of detached leaf discs exposed to about 2000 μmol m−1 s−1 photon flux density (PFD) for 2 h under controlled conditions (25 to 28 °C, fully hydrated). Differences in susceptibility to photodamage among species were not significant in the open and 10% full sun treatments. But in 5% full sun, B. platyphylla sustained a significantly greater photodamage than other species, probably associated with having the lowest photosynthetic capacity indicated by light‐saturated photosynthetic rate (B. platyphylla, 9·87, 5·85 and 2·82; Q. mongolica, 8·05, 6·28 and 4·41; A. mono, 7·93, 6·11 and 5·08 μmol CO2 m−1 s−1for open, 10% and 5% full sun, respectively). To simulate a gap formation and assess its complex effects including high temperature and water stress in addition to strong light on the susceptibility to photoinhibition, we examined photoinhibition in the field by means of monitoring ΔF/F′m on the first day of transfer to natural daylight. Compared with ΔF/F′m in AM, the lower ΔF/F′m in PM responding to lower PFD following high PFD around noon indicated that photoinhibition occurred in plants grown in 10 and 5% full sun. The diurnal changes of ΔF/F′m showed that Q. mongolica grown in 5% full sun was less susceptible to photoinhibition than A. mono although they showed little differences both in photosynthetic capacity in intact leaves and susceptibility to photoinhibition based on leaf disc measurements. These results suggest that shade‐grown Q. mongolica had a higher tolerance for additional stresses such as high temperature and water stress in the field, possibly due to their lower plasticity in leaf anatomy to low light environment.
Cell growth analysis during steady and non‐steady growth in leaves of perennial ryegrass (Lolium perenne L.) subject to defoliation
Authors: Schäufele R. , Schnyder H.
Published: March 2003

The effect of defoliation on leaf elongation rate (LER) and on the spatial distribution of epidermal cell lengths in the leaf growth zone was studied in vegetative main tillers of perennial ryegrass (Lolium perenne L. cv Modus) grown in a controlled environment. A new material approach was used to analyse the responses of epidermal cell expansion and production during the initial, non‐steady growth phase following defoliation. The analysis involved assigning an identity to individual expanding cells, assessing the displacement and estimating the expansion of cells with assigned identity during day 1 and day 2 after defoliation. LER decreased by 34% during the first 2 d after defoliation and did not recover to the pre‐defoliation rate within the 14 day regrowth period. Decreased LER on day 1 and day 2 after defoliation was associated with (i) a decrease in the length of the leaf growth zone; (ii) a decrease in the length at which epidermal cells stopped expanding; (iii) a reduced expansion of cells at intermediate growth stages; and (iv) a reduction in cell production (i.e. division) and an associated decrease in the number of expanding cells in the growth zone. However, defoliation had no effect on the expansion of cells located in the proximal part of the growth zone. Reduced LER at 14 d after defoliation was associated with a reduced cell production rate (27% lower than the pre‐defoliation rate) and decreased final cell size (− 28%).
Evidence implicating a novel thiol methyltransferase in the detoxification of glucosinolate hydrolysis products in Brassica oleracea L.
Authors: Attieh J. , Kleppinger‐Sparace K. F. , Nunes C. , Sparace S. A. , Saini H. S.
Published: March 2003

The physiological relevance of a novel thiol methyltransferase from cabbage, and its possible role in sulphur metabolism have been investigated. The enzyme was absent from the chloroplast, the site of sulphate reduction, and was localized in the cytosol. Potential substrates were initially screened on the basis of their ability to inhibit the methylation of iodide, a previously known substrate for the enzyme. Thiocyanate, 4,4′‐thiobisbenzenethiol, thiophenol, and thiosalicylic acid were identified as possible substrates. Methylation of these thiols by the purified enzyme using [Methyl‐3H]S‐adenosyl‐
L‐methionine confirmed their nature as substrates. The purified enzyme strongly preferred thiocyanate as a methyl acceptor. The enzyme had Km values of 11, 51, 250 and 746 mmol m−3 for thiocyanate, 4,4′‐thiobisbenzenethiol, thiophenol and thiosalicylic acid, respectively. The identity of methylthiocyanate as the product of thiocyanate methylation by the purified enzyme was confirmed by mass spectrometry. The enzyme was strictly associated with glucosinolate‐containing plants. Thiol substrates of the enzyme are known products of glucosinolate hydrolysis. Our observations indicate that this enzyme could be involved in the detoxification of reactive thiols produced upon glucosinolate degradation in these plants.
Detopping causes production of intercellular space occlusions in both the cortex and infected region of soybean nodules
Authors: James E. K. , Iannetta P. P. M. , Deeks L. , Sprent J. I. , Minchin F. R.
Published: March 2003

A structural analysis was conducted to determine whether glycoprotein‐containing intercellular space occlusions are involved in medium‐term regulation of O2 diffusion in soybean (Glycine max) nodules. Alterations in O2 diffusion were induced by a 3 h detopping treatment, and glycoprotein was immunolocalized with the monoclonal antibodies MAC236 and MAC265. Western blots of unstressed nodules revealed that these antibodies recognize antigens with two different molecular weights in soybean nodules. Tissue printing of halved nodules showed that both antigens were present in fresh nodules from control and 3 h detopped plants. The main localization appeared to be the inner cortex, but some immunolabelling also occurred in the infected region. ELISAs demonstrated a significant increase in total nodule concentration of intercellular glycoprotein following detopping, and cryosections of fresh nodules from this treatment also showed localization of antigens within the intercellular spaces of the infected region. The production of intercellular space occlusions in both the mid‐cortex and infected regions after 3 h detopping was confirmed by light microscopy and silver‐enhanced immunolabelling; cortical changes were quantified by image analysis techniques. Electron microscopy revealed that the occlusions within the infected region were less dense and less heavily labelled than those in the cortex. These results are discussed in relation to O2 diffusion regulation in soybean nodules
Why are higher plants green? Evolution of the higher plant photosynthetic pigment complement
Authors: Nishio J. N.
Published: March 2003

The physiological reason that higher plants are green is unknown. Other photosynthetic organisms utilize pigments that strongly absorb green light; therefore, there must have been natural forces that ‘selected’ the photosynthetic pigments found in higher plants. Based on previously published data and our recent findings about green light and photosynthesis within leaves (Sun et al.), a specific functional role is described for the primary photosynthetic pigments of higher plants, that were derived from green algal progenitors. The particular absorptive characteristics of chlorophylls a and b appear to perform two contradictory, but necessary functions in higher plants. Firstly, chlorophylls a and b absorb light for maximum utilization under non‐saturating conditions, a function that is well understood. Secondly, they can act as protective pigments under over‐saturating light conditions, when absorbed light is dissipated as heat. Under such conditions, a significant portion of light can also be efficiently utilized, especially in the bottom portion of the leaf, that is mainly illuminated by green light and not down‐regulated. The second function may have been the selective force that gave rise to the extremely successful terrestrial plants, that evolved from green algae.
Source metabolism dominates the control of source to sink carbon flux in tuberizing potato plants throughout the diurnal cycle and under a range of environmental conditions
Authors: Sweetlove L. J. , Hill S. A.
Published: March 2003

The aim of this work was to investigate the effect of environmental conditions on the control of source to sink carbon flux in potato during tuberization. Top down metabolic control analysis was used to quantitatively determine the extent to which source and sink reactions control the flux from CO2 to the tuber. Under all conditions investigated (beginning of the dark period, low nitrogen availability, variation in temperature) source metabolism accounts for at least 80% of the control. In plants grown with limiting nitrogen, or when measurements were made at low temperature the extent to which source metabolism controls flux increased to 90%.
Influence of carbon dioxide enrichment, ozone and nitrogen fertilization on cotton (Gossypium hirsutumL.) leaf and root composition
Authors: Booker F. L.
Published: March 2003

The objective of this study was to test whether elevated [CO2], [O3] and nitrogen (N) fertility altered leaf mass per area (LMPA), non‐structural carbohydrate (TNC), N, lignin (LTGA) and proanthocyanidin (PA) concentrations in cotton (Gossypium hirsutumL.) leaves and roots. Cotton was grown in 14 dm3pots with either sufficient (0·8 g N dm−3) or deficient (0·4 and 0·2 g N dm−3) N fertilization, and treated in open‐top chambers with either ambient or elevated (+175 and+350μmol mol−1) [CO2] in combination with either charcoal‐filtered air (CF) or non‐filtered air plus 1·5 times ambient [O3]. At about 50 d after planting, LMPA, starch and PA concentrations in canopy leaves were as much as 51–72% higher in plants treated with elevated [CO2] compared with plants treated with ambient [CO2], whereas leaf N concentration was 29% lower in elevated [CO2]‐treated plants compared with controls. None of the treatments had a major effect on LTGA concentrations on a TNC‐free mass basis. LMPA and starch levels were up to 48% lower in plants treated with elevated [O3] and ambient [CO2] compared with CF controls, although the elevated [O3] effect was diminished when plants were treated concurrently with elevated [CO2]. On a total mass basis, leaf N and PA concentrations were higher in samples treated with elevated [O3] in ambient [CO2], but the difference was much reduced by elevated [CO2]. On a TNC‐free basis, however, elevated [O3] had little effect on tissue N and PA concentrations. Fertilization treatments resulted in higher PA and lower N concentrations in tissues from the deficient N fertility treatments. The experiment showed that suppression by elevated [O3] of LMPA and starch was largely prevented by elevated [CO2], and that interpretation of [CO2] and [O3] effects should include comparisons on a TNC‐free basis. Overall, the experiment indicated that allocation to starch and PA may be related to how environmental factors affect source–sink relationships in plants, although the effects of elevated [O3] on secondary metabolites differed in this respect.
Molecular markers for ozone stress isolated by suppression subtractive hybridization: specificity of gene expression and identification of a novel stress‐regulated gene
Authors: Sävenstrand H. , Brosché M. , Ängehagen M. , Strid Å.
Published: March 2003

Suppression subtractive hybridization was used to identify genes regulated by ozone (100 nmol mol−1) in Pisum sativum. One novel gene (named PsUod1) was found. In addition, mRNA levels for four genes (encoding lipid transfer protein, pre‐hevein‐like protein, leucine‐rich repeat protein, and disease‐resistance response protein 230), which previously were shown to be regulated by biotic stress, increased. Finally, mRNA species for two genes (encoding extensin and pathogenesis‐related protein 4A), previously shown to be regulated by ozone in other species, were found to increase in abundance. The ozone‐specificity of the expression of these genes was studied by using UV‐B radiation. PsUod1 and the genes encoding extensin, leucine‐rich repeat protein, and disease‐resistance response protein 230, were differentially regulated when comparing ozone and UV‐B. Moreover, the mRNA levels for extensin, leucine‐rich repeat protein and disease‐resistance response protein 230 all increased under NaCl and aluminium stress and after wounding, whereas the message abundance for PsUod1 was unchanged under these stresses. Thus, in general, ozone caused changes similar to wounding, salt stress and aluminium stress, whereas UV‐B radiation regulated gene expression differently.
Strategies providing success in a variable habitat: I. Relationships of environmental factors and dominance of Cladophora glomerata
Authors: Ensminger I. , Hagen C. , Braune W.
Published: March 2003

Dominance of macrophytes and their response to environmental factors were studied in the river Ilm, Thuringia/ Germany with special reference to Cladophora glomerata (L.) Kütz. Macroalgae showed a growth peak in spring with C. glomerata, Ulothrix zonata Kütz., Lemanea fluviatilis (L.) C. Agardh and Audouinella sp. being the dominant species. Shortly after this peak, a rapid decline of macrophyte substrate coverage was observed. Only C. glomerata revealed a second growth peak in late summer/early autumn. Frequent disturbances of the macrophyte assemblage by floods resulted repeatedly in an almost complete wash out of benthic organisms. After summer floods C. glomerata was the species that recolonized the substrate. At high‐light sites, faster recovery of C. glomerata was observed as compared to low‐light sites. This is discussed in relation to the life cycle of C. glomerata. Among the physical and chemical parameters that were analysed, irradiance, current velocity, pH, soluble reactive phosphorus and ammonia‐nitrogen accounted for most of the observed patterns of dominance of C. glomerata.
Photosynthesis and conductance of spring wheat ears: field response to free‐air CO2 enrichment and limitations in water and nitrogen supply
Authors: Wechsung F. , Garcia R. L. , Wall G. W. , Kartschall T. , Kimball B. A. , Michaelis P. , Jr P. J. Pinter , Wechsung G. , Grossman‐Clarke S. , Lamorte R. L. , Adamsen F. J. , Leavitt S. W. , Thompson T. L. , Matthias A. D. , Brooks T. J.
Published: March 2003

The mid‐day responses of wheat ear CO2 and water vapour exchange to full‐season CO2 enrichment were investigated using a Free‐Air CO2 Enrichment (FACE) apparatus. Spring wheat [Triticum aestivum (L). cv. Yecora Rojo] was grown in two experiments under ambient and elevated atmospheric CO2 (Ca) concentrations (approximately 370 μmol mol−1 and 550 μmol mol−1, respectively) combined first with two irrigation (Irr) schemes (Wet: 100% and Dry: 50% replacement of evapotranspiration) and then with two levels of nitrogen (N) fertilization (High: 350, Low: 70 kg ha−1 N). Blowers were used for Ca enrichment. Ambient Ca plots were exposed to blower induced winds as well the Ca× N but not in the Ca× Irr experiment. The net photosynthesis for the ears was increased by 58% and stomatal conductance (gs) was decreased by 26% due to elevated Ca under ample water and N supply when blowers were applied to both Ca treatments. The use of blowers in the Ca‐enriched plots only during the Ca× Irr experiment (blower effect) and Low N supply restricted the enhancement of net photosynthesis of the ear due to higher Ca. In the latter case, the increase of net photosynthesis of the ear amounted to 26%. The decrease in gs caused by higher Ca was not affected by the blower effect and N treatment. The mid‐day enhancement of net photosynthesis due to elevated Ca was higher for ears than for flag leaves and this effect was most pronounced under ample water and N supply. The contribution of ears to grain filling is therefore likely to increase in higher Ca environments in the future. In the comparison between Wet and Dry, the higher Ca did not alter the response of net photosynthesis of the ear and gs to Irr. However, Ca enrichment increased the drought tolerance of net photosynthesis of the glume and delayed the increase of the awn portion of net photosynthesis of the ear during drought. Therefore, the role of awns for maintaining high net photosynthesis of the ear under drought may decrease when Ca increases.
Lactate metabolism in potato tubers deficient in lactate dehydrogenase activity
Authors: Sweetlove L. J. , Dunford R. , Ratcliffe R. G. , Kruger N. J.
Published: March 2003

The aim of this work was to investigate the effect of decreased activity of lactate dehydrogenase (EC 1.1.1.27; LDH) on lactate metabolism in potato tubers. By expressing a cDNA‐encoding potato tuber LDH in the antisense orientation, we generated transgenic potato plants with a preferential decrease in two of the five isozymes of LDH. Surprisingly, transgenic tubers grown under normoxic conditions did not contain less lactate, but rather instead contained approximately two‐fold more lactate than control tubers. This result is explicable if the decreased isozymes of LDH are responsible for the oxidation of lactate to pyruvate in vivo. This was confirmed by measurements of the rate of metabolism of lactate supplied to tuber discs: the rate in transgenic tubers was approximately half that of control tubers. The decrease in LDH activity had no measurable effect on the accumulation of lactate in cold‐stored tubers under anoxia, nor during the subsequent utilization of this lactate upon return to normoxia. In both control and transgenic tubers, the accumulation of lactate during anoxia was not accompanied by an induction of LDH activity or a change in isozyme distribution. In contrast, the metabolism of lactate after a period of anoxia was accompanied by a two‐fold increase in LDH activity and the induction of two isozymes that were distinct from those which had been decreased in the transgenic plants.
Elevated CO2 stimulates associative N2 fixation in a C3 plant of the Chesapeake Bay wetland
Authors: Dakora F. D. , Drake B. G.
Published: March 2003

In this study, the response of N2 fixation to elevated CO2 was measured in Scirpus olneyi, a C3 sedge, and Spartina patens, a C4 grass, using acetylene reduction assay and 15N2 gas feeding. Field plants grown in PVC tubes (25 cm long, 10 cm internal diameter) were used. Exposure to elevated CO2 significantly (P< 0·05) caused a 35% increase in nitrogenase activity and 73% increase in 15N incorporated by Scirpus olneyi. In Spartina patens, elevated CO2 (660 ± 1 μmol mol−1) increased nitrogenase activity and 15N incorporation by 13 and 23%, respectively. Estimates showed that the rate of N2 fixation in Scirpus olneyi under elevated CO2 was 611 ± 75 ng 15N fixed plant−1 h−1 compared with 367 ± 46 ng 15N fixed plant−1 h−1 in ambient CO2 plants. In Spartina patens, however, the rate of N2 fixation was 12·5 ± 1·1 versus 9·8 ± 1·3 ng 15N fixed plant−1 h−1 for elevated and ambient CO2, respectively. Heterotrophic non‐symbiotic N2 fixation in plant‐free marsh sediment also increased significantly (P< 0·05) with elevated CO2. The proportional increase in 15N2 fixation correlated with the relative stimulation of photosynthesis, in that N2 fixation was high in the C3 plant in which photosynthesis was also high, and lower in the C4 plant in which photosynthesis was relatively less stimulated by growth in elevated CO2. These results are consistent with the hypothesis that carbon fixation in C3 species, stimulated by rising CO2, is likely to provide additional carbon to endophytic and below‐ground microbial processes.
Hydraulic architecture and water flow in growing grass tillers (Festuca arundinacea Schreb.)
Authors: Martre P. , Cochard H. , Durand J.‐L.
Published: March 2003

The water relations and hydraulic architecture of growing grass tillers (Festuca arundinacea Schreb.) are reported. Evaporative flux density, E (mmol s−1 m−2), of individual leaf blades was measured gravimetrically by covering or excision of entire leaf blades. Values of E were similar for mature and elongating leaf blades, averaging 2·4 mmol s−1 m−2. Measured axial hydraulic conductivity, Kh (mmol s−1 mm MPa−1), of excised leaf segments was three times lower than theoretical hydraulic conductivity (Kt) calculated using the Poiseuille equation and measurements of vessel number and diameter. Kt was corrected (Kt*) to account for the discrepancy between Kh and Kt and for immature xylem in the basal expanding region of elongating leaves. From base to tip of mature leaves the pattern of Kt* was bell‐shaped with a maximum near the sheath–blade joint (≈ 19 mmol s−1 mm MPa−1). In elongating leaves, immature xylem in the basal growing region led to a much lower Kt*. As the first metaxylem matured, Kt* increased by 10‐fold. The hydraulic conductances of the whole root system, (mmol s−1 MPa−1) and leaf blades, (mmol s−1 MPa−1) were measured by a vacuum induced water flow technique. and were linearly related to the leaf area downstream. Approximately 65% of the resistance to water flow within the plant resided in the leaf blade. An electric‐analogue computer model was used to calculate the leaf blade area‐specific radial hydraulic conductivity, (mmol s−1 m−2 MPa−1), using , Kt* and water flux values. values decreased with leaf age, from 21·2 mmol s−1 m−2 MPa−1 in rapidly elongating leaf to 7·2 mmol s−1 m−2 MPa−1 in mature leaf. Comparison of and values showed that ≈ 90% of the resistance to water flow within the blades resided in the liquid extra‐vascular path. The same algorithm was then used to compute the xylem and extravascular water potential drop along the liquid water path in the plant under steady state conditions. Predicted and measured water potentials matched well. The hydraulic design of the mature leaf resulted in low and quite constant xylem water potential gradient (≈ 0·3 MPa m−1) throughout the plant. Much of the water potential drop within mature leaves occurred within a tenth of millimetre in the blade, between the xylem vessels and the site of water evaporation within the mesophyll. In elongating leaves, the low Kt* in the basal growth zone dramatically increased the local xylem water potential gradient (≈ 2·0 MPa m−1) there. In the leaf elongation zone the growth‐induced water potential difference was ≈ 0·2 MPa.
Recent advances in the role and biosynthesis of ascorbic acid in plants
Authors: Conklin P. L.
Published: March 2003

The past few years have provided many advances in the role and biosynthesis of L‐ascorbic acid (AsA) in plants. There is an increasing body of evidence confirming that AsA plays an important role in the detoxification of reactive oxygen species. The role of AsA in photoprotection has been confirmed in vivo with the use of Arabidopsis mutants. A player in the defence against reactive oxygen species, AsA peroxidase, has been extensively studied at the molecular level, and regulation of this key enzymatic activity appears to occur at several levels. As a cofactor in the hydroxylation of prolyl and lysl‐residues by peptidyl‐prolyl and ‐lysyl hydroxylases, AsA plays a part in cell wall synthesis, defence, and possibly cell division. The maintenance of reduced levels of AsA appears to be highly regulated, involving the interplay of both monodehydroascorbate and dehydroascorbate reductases and possibly auxin. A major breakthrough in plant AsA biosynthesis has been made recently, and strong biochemical and genetic evidence suggest that GDP‐mannose and L‐galactose are key substrates. In addition, evidence for an alternative AsA biosynthetic pathway(s) exists and awaits additional scrutiny. Finally, newly described Arabidopsis mutants deficient in AsA will further increase our understanding of AsA biosynthesis
The distribution of xylem hydraulic resistance in the fruiting truss of tomato
Authors: Malone M. , Andrews J.
Published: March 2003

The distribution of hydraulic resistances in xylem throughout the pathway leading to the tomato fruit was investigated. Previous work had indicated that there were large resistances within the supporting sections of this pathway (the peduncle and pedicel), perhaps associated with interruptions in the xylem. These high resistances are believed to impede calcium flux into the fruit and thus impair fruit development. It is shown here that fruit on intact plants do not shrink detectably during drought, even when the drought is sufficient to cause marked shrinkage of leaves and visible wilting of the shoot. In explants, it is possible to induce back‐flow from the fruit into the stem (probably via the xylem) but this flow is small and very slow. These observations support the view that there is a large hydraulic resistance in the pathway between fruit and stem. When pulses of water were made available within explants, by scorching of one leaflet, there was a rapid swelling of leaves and sepals. Such rapid fluxes indicate the presence of strong hydraulic (xylem) connections throughout the pathway between leaf and calyx. This shows that there are no significant hydraulic constrictions in the xylem proximal to the calyx. This finding is contrary to some previous conclusions but it is supported by experiments with dyes which showed continuous, functional xylem throughout the peduncle and pedicel. Calculations show that over 90% of the hydraulic resistance between stem and fruit must reside within the fruit pericarp. Implications for calcium nutrition are discussed.
Influence of atmospheric oxygen on leaf structure and starch deposition in Arabidopsis thaliana
Authors: Ramonell K. M. , Kuang A. , Porterfield D. M. , Crispi M. L. , Xiao Y. , Mcclure G. , Musgrave M. E.
Published: March 2003

Plant culture in oxygen concentrations below ambient is known to stimulate vegetative growth, but apart from reports on increased leaf number and weight, little is known about development at subambient oxygen concentrations. Arabidopsis thaliana (L.) Heynh. (cv. Columbia) plants were grown full term in pre‐mixed atmospheres with oxygen partial pressures of 2·5, 5·1, 10·1, 16·2, and 21·3 kPa O2, 0·035 kPa CO2 and the balance nitrogen under continuous light. Fully expanded leaves were harvested and processed for light and transmission electron microscopy or for starch quantification. Growth in subambient oxygen concentrations caused changes in leaf anatomy (increased thickness, stomatal density and starch content) that have also been described for plants grown under carbon dioxide enrichment. However, at the lowest oxygen treatment (2·5 kPa), developmental changes occurred that could not be explained by changes in carbon budget caused by suppressed photorespiration, resulting in very thick leaves and a dwarf morphology. This study establishes the leaf parameters that change during growth under low O2, and identifies the lower concentration at which O2 limitation on transport and biosynthetic pathways detrimentally affects leaf development.
UV‐B and UV‐C induction of NADP‐malic enzyme in tissues of different cultivars of Phaseolus vulgaris (bean)
Authors: Casati P. , Andreo C. S.
Published: March 2003

The effects of the treatment of different tissues of three bean cultivars (Pinto, Vilmorin and Arroz) with ultra‐violet (UV) UV‐B and UV‐C radiation and red light on the activity, quantity and RNA levels of NADP‐malic enzyme (NADP‐ME) were determined. Exposure to UV‐B radiation for 8 h caused a marked increase of NADP‐ME from leaves, stems and roots in the three cultivars studied. A similar induction was observed in the leaves and stems after 8 h of exposure under UV‐C, but not in the roots, suggesting that a different signal might be acting to induce the expression of NADP‐ME after UV‐B and UV‐C exposure. In contrast, red light was ineffective in inducing NADP‐ME in either tissue, so the regulation of the expression of this enzyme is phytocrome‐independent. The activity of superoxide dismutase, ascorbate peroxidase, catalase and peroxidase was also different in plants treated with UV‐B, UV‐C and photosynthetically active radiation, suggesting that various pathways may be acting in the regulation of these enzymes by UV‐B and UV‐C. Reactive oxygen species (ROS) were also required for UV‐B induction of NADP‐ME, as the addition of ascorbic acid before UV‐B treatment prevented NADP‐ME induction, whereas salicylic acid was not effective in inducing the enzyme, showing that NADP‐ME induction by UV‐B is ROS dependent but salicylic acid independent.
Antioxidant activities of red versus green leaves in Elatostema rugosum
Authors: Neill S. O. , Gould K. S. , Kilmartin P. A. , Mitchell K. A. , Markham K. R.
Published: March 2003

Anthocyanin biosynthesis in leaves increases under stresses which also generate reactive oxygen species (ROS). In the present study the hypothesis that red leaves are better equipped to scavenge ROS than green leaves was tested. Antioxidants in leaf extracts from red and green morphs of Elatostema rugosum were identified, and activities quantified using enzymatic and α,α‐diphenyl‐β‐picrylhydrazyl (DPPH) assays and cyclic voltammetry. Red leaves from E. rugosum held greater amounts of superoxide dismutase, catalase, anthocyanins, and hydroxycinnamic acids, were significantly more effective at scavenging DPPH radicals, and produced higher voltammetric currents than green leaves. Anthocyanins contributed to the antioxidant pool more than all other constituent phenolics. Anthocyanin concentrations, and antioxidant activities declined with leaf age. Purified anthocyanin fractions displayed oxidative activities at both pH 7·0 and pH 5·5. Implications of the antioxidant potential of anthocyanin in its cytoplasmic and vacuolar locations are discussed.
Activation of an oxidative burst is a general feature of sensitive plants exposed to the air pollutant ozone
Authors: Wohlgemuth H. , Mittelstrass K. , Kschieschan S. , Bender J. , Weigel H.‐J. , Overmyer K. , Kangasjärvi J. , Sandermann H. , Langebartels C.
Published: March 2003

Ozone exposure stimulates an oxidative burst in leaves of sensitive plants, resulting in the generation and accumulation of hydrogen peroxide (H2O2) in tobacco and tomato, and superoxide (O2–•) together with H2O2 in Arabidopsis accessions. Accumulation of these reactive oxygen species (ROS) preceded the induction of cell death, and both responses co‐occurred spatially in the periveinal regions of the leaves. Re‐current ozone exposure of the sensitive tobacco cv. Bel W3 in closed chambers or in the field led to an enlargement of existing lesions by priming the border cells for H2O2 accumulation. Open top chamber experiments with native herbaceous plants in the field showed that Malva sylvestris L. accumulates O2–• at those sites that later exhibit plant cell death. Blocking of ROS accumulation markedly reduced ozone‐induced cell death in tomato, Arabidopsis and M. sylvestris. It is concluded that ozone triggers an in planta generation and accumulation of H2O2 and/or O2–• depending on the species, accession and cultivar, and that both these reactive oxygen species are involved in the induction of cell death in sensitive crop and native plants.
Induction of the Na+/Pi cotransport system in the plasma membrane of Chara corallina requires external Na+ and low levels of Pi
Authors: Mimura T. , Reid R. J. , Ohsumi Y. , Smith F. A.
Published: March 2003

It was shown in previous studies that the giant freshwater alga Chara corallina does not control its Na+‐dependent Pi uptake by monitoring the internal Pi concentration and it was hypothesized that Chara may instead detect changes in Pi supply from the environment. The present work investigated the conditions that control the induction and inactivation of high affinity Na+/Pi influx in Chara. Withdrawal of Pi from the external medium resulted in a gradual increase in the rate of uptake measured immediately after Pi was resupplied. The increase continued for at least 7 d of starvation. In the initial stages, 0·5 or 1 µm Pi were more effective at inducing transport activity than no Pi, suggesting that low levels of Pi are actually required for induction. The high Na+‐dependent Pi uptake observed in Pi‐starved cells was inactivated by treatment with as little as 1 µm Pi over 6 d. External Na+ plays a major role in controlling the capacity for Na+/Pi cotransport activity, and in the absence of Na+, both induction and inactivation were either delayed or abolished. Na+ starvation stimulated Na+ uptake even though there were no measurable changes in the concentrations of Na+, or of K+ or Pi in either the vacuole or cytoplasm. It was concluded that both substrate (Pi) and driver ion (Na+) are required at adequate concentrations for the induction of the cotransporter. In the case of Pi, it was suggested that passive leakage of Pi from the cell into the apoplast is sufficient for this purpose but that supplementation by up to 1 µm Pi is more effective at the earlier stage. A mechanism for sensing the external supply of Pi is proposed.
Environmental and metaboliccontrol of LHCII protein phosphorylation: revealing the mechanismsfor dual regulation of the LHCII kinase
Authors: Hou C.‐X. , Pursiheimo S. , Rintamäki E. , Aro E.‐M.
Published: March 2003

Light‐harvesting complex II (LHCII) protein phosphorylation inplant chloroplasts is under complex regulation. Combination of the invivo monitoring of LHCII protein phosphorylation (by immunoblotting)with the in vitro[γ‐32P]ATPphosphorylation assays revealed that the basic activation/deactivationmodel of the LHCII kinase, regulated by reversible occupation/releaseof plastoquinol at the plastoquinol oxidation (Qo) siteof the cytochrome b6f (cyt b6f) complex, isconsistent with, but not sufficient to explain the data obtainedwith isolated chloroplasts, leaf discs or intact leaves. Not onlythe light conditions but also the metabolic state of the entireplant, particularly the sugar metabolism, exerted a control overLHCII protein phosphorylation. Feeding of leaves with glucose (alsowith glutathione) activated the LHCII kinase in darkness. On the otherhand, independently of the basic activation/deactivationmechanism of the kinase, a strong inhibition of LHCII protein phosphorylationoccurred in vivo at increasing irradiances and even at lowlight conditions, depending on the metabolic state of the plant.Both the experiments with intact chloroplasts and the reconstitutionexperiments with isolated thylakoids to mimic LHCII kinase inhibition,disclosed that the kinase in its activated state (plastoquinol at theQo site of cyt b6f complex) is protected againstinhibition by thiol reductants. However, directly upon deactivationof the kinase (release of plastoquinol from the Qo site) itbecomes a target for inhibition by thiol reductants. Thus the twointerdependent regulatory systems of the LHCII kinase, the constantlyoccurring activation and deactivation on the one hand and the inhibitionby thiol reductants on the other, are strongly dependent on theconcentration of reducing equivalents in the chloroplast stroma.A scheme demonstrating the interconversion of activated, deactivated andinhibited states of the LHCII kinase in the chloroplast environmentof intact leaves is presented.
Genetic variation among field isolates of Pyrenopeziza brassicae
Authors: Majer D. , Lewis B. G. , Mithen R.
Published: March 2003

Amplified fragment length polymorphism (AFLP) markers were used to determine genetic diversity and population structure of Pyrenopeziza brassicae, the causal agent of light leaf spot of Brassica spp. Fungal isolates were sampled from six regions in the UK, one region in Germany and one region in France. A high level of genetic diversity was found (HT = 58%), with most variation attributed to within regions (HS = 43%), which suggests that sexual reproduction is frequent. FST values suggested significant population differentiation between England and the continent, but not between Scotland and England and Scotland and the continent. Overall, a moderate but significant level of regional differentiation was found (FST = 16 ± 4.0). There was no correlation between FST values and distance, indicating that long‐distance dispersal by natural factors does not occur at high frequencies. However, the lack of differentiation among populations from Aberdeen, Winchester and Cambridge suggests that seed transmission or other artificial methods of dispersal may be important.
The use of species‐specific PCR‐based assays to analyse Fusarium ear blight of wheat
Authors: Doohan F. M. , Parry D. W. , Jenkinson P. , Nicholson P.
Published: March 2003

Polymerase chain reaction (PCR) assays for the detection of various Fusarium species and Microdochium nivale subspecies were compared with conventional visual disease assessment using a field plot of wheat in which the central subplot was inoculated with F. culmorum. Visual disease assessment was performed on a range of samples taken from each of 15 subplots at growth stage 80. At harvest, each sample was divided into its component parts, i.e. grain, glume and rachis, and species‐specific PCR analysis was used to detect the presence of F. culmorum, F. poae, F. avenaceum, F. graminearum, M. nivale var. majus and M. nivale var. nivale. Within the inoculated subplot there was good correlation between visual disease assessment and PCR analysis, both techniques indicating a high incidence of F. culmorum in this region. According to the visual disease assessment results, there was also a relatively high incidence of F. culmorum in most other regions of the field plot. However, according to PCR analysis the incidence of F. culmorum in many of the other subplots was relatively low and F. poae, M. nivale var. majus and var. nivale, and F. avenaceum were detected within the grain, glume and rachis tissues of many of the ear samples from these subplots. F. poae predominated in the glume component of ears and M. nivale var. majus and var. nivale in the rachis component. M. nivale PCR results revealed that 64% of infected samples involved var. majus, and 36% var. nivale. PCR analysis has highlighted some difficulties that may arise when using visual assessment for studying disease complexes.
Incidence of weed reservoirs and vectors of tomato spotted wilt tospovirus on southern Tasmanian lettuce farms
Authors: Wilson C. R.
Published: March 2003

Thrips species and tomato spotted wilt virus (TSWV) alternate weed hosts were surveyed on two lettuce farms in southern Tasmania during 1994 and 1995. Only one known vector species, Thrips tabaci, was found at either site, comprising on average 36.8% of the total monthly catch. A major peak of thrips activity in the summer corresponded with an increase of disease in autumn harvested lettuce. Two thrips species new for Tasmania were recorded, Pseudanaphothrips achaetus and Tenothrips frici. Infection patterns within the crop indicated that localized weed infestations were the most likely reservoir of virus. ELISA testing showed that TSWV was present in a range of dicotyledonous weed species, although usually infecting only a low percentage of the plants. Arctotheca calendula appeared to be the single most important reservoir host species at one property, whilst this species and Sonchus oleraceus, Malva sylvestris, Brassica rapa ssp. silvestris, Erodium moschatum and Trifolium sp. were probably the most important reservoirs at the other property. Two new natural TSWV host species were recorded, Erodium moschatum and Brassica rapa ssp. silvestris. The property with the highest incidence of TSWV‐infected lettuce had a relatively higher proportion of virus‐infected weeds but less thrips activity during the infection period.
Combining proteolytic and phloroglucinol‐producing bacteria for improved biocontrol of Pythium‐mediated damping‐off of sugar beet
Authors: Dunne C. , Moënne‐Loccoz Y. , McCarthy J. , Higgins P. , Powell J. , Dowling D. N. , O'Gara F.
Published: March 2003

Pseudomonas fluorescens F113 and Stenotrophomonas maltophilia W81 protect sugar beet from Pythium‐mediated damping‐off through production of the antifungal secondary metabolite 2,4‐diacetylphloroglucinol and extracellular proteolytic activity, respectively. In this study, the two biocontrol strains were combined in a consortium, with the objective of improving upon the level of protection achieved when using each strain singly. Growth and in vitro production of 2,4‐diacetylphloroglucinol by F113 and extracellular lytic enzymes by W81 were not affected when inoculated in combination. The abilities of W81 and F113 to colonize the rhizosphere of sugar beet were essentially similar when the two strains were applied singly or coinoculated onto seeds in a 1 : 1 ratio, both in natural soil microcosms and under field conditions. Concomitantly, single inoculation with W81 or F113 effectively prevented colonization of sugar beet seeds by Pythium spp. in soil microcosms, without the necessity for combining both strains. However, this parity was not reflected in seed emergence where the combination of W81 and F113 significantly enhanced final sugar beet stands (to the level achieved with chemical pesticides) under microcosm conditions at 28 days after sowing. In a field experiment, the only inoculation treatment capable of conferring effective protection of sugar beet was that in which W81 and F113 were coinoculated, and this treatment proved equivalent to the use of chemical fungicides. In conclusion, when compared with single inoculations of either biocontrol strain, the combined use of a phloroglucinol‐producing P. fluorescens and a proteolytic S. maltophilia improved protection of sugar beet against Pythium‐mediated damping‐off.
Determination of Ralstonia (Pseudomonas) solanacearum rDNA subgroups by PCR tests
Authors: Seal  , Taghavi  , Fegan  , Hayward  , Fegan
Published: March 2003

Rapid and sensitive polymerase chain reaction (PCR) methods are described for determination of the two 16 S rDNA subgroups of Ralstonia solanacearum, the causal agent of bacterial wilt. A third subgroup consisting of Indonesian R. solanacearum isolates belonging to Division II, the blood disease bacterium and Pseudomonas syzygii can also be identified. Primers were designed to sequences within R. solanacearum 16 S rDNA (equivalent to Escherichia coli 16 S rDNA positions 74–97, 455–475, 1454–1474), and the internal transcribed spacer region between the 16 S and 23 S rDNA genes. Different combinations of forward and reverse primers allowed selective PCR amplification of (a) R. solanacearum Division I (biovars 3, 4 and 5), (b) Division II (biovars 1, N2, and 2) including the blood disease bacterium and P. syzygii, or (c) amplification of Division II only except for five biovar 1, 2 or N2 isolates of R. solanacearum from Indonesia, P. syzygii and the BDB. A total of 104 R. solanacearum, 14 blood disease bacterium and 10 P. syzygii isolates were tested. Simultaneous detection of species and subdivision was achieved by designing a multiplex PCR test in which a 288‐base pair (bp) band is produced by all R. solanacearum isolates, and an additional 409‐bp band in Division I strains.