The effect of low temperature on patterns of cell division in developing second leaves of wild-type and slender mutant barley (Hordeum vulgare L.)

This study reports on investigations into the effect of long-term growth at reduced temperatures on cell elongation and cell division in the wild type and a temperature-insensitive ( slender ) mutant of barley. Plants were grown under two temperature regimes (20 and 5 °C) and the mitotic index, cell doubling time and cell lengths over the division and elongation zone were monitored at several stages of development in the second leaf. Leaf length and leaf growth rates were characteristically greater in the slender mutant than in the wild type and this was greatly exaggerated by growth at low temperature. Cell length and the length of the division zone were also greater in the slender mutant than in the wild type, and growing the plants at reduced temperature (5 °C) shortened cell lengths only in the wild type. The slender mutant had a higher mitotic index than the wild type, although in neither genotype was change in the mitotic index observed following growth at reduced temperature. Cell doubling time, on the other hand, was reduced by growth at reduced temperature in the wild type but not in the slender mutant. Thus, the data suggest very different growth responses to low temperature in the two genotypes. The results are discussed in terms of the ability of plants to sense their environment and optimize their metabolism for future growth.     

Time-dependent changes in polypeptide and translatable mRNA levels caused by NaCl in barley roots

The effect of salt stress on polypeptide and translatable mRNA levels was examined in roots of barley ( Hordeum vulgare L. cv. CM 72). A salt-shock time course (200 m M NaCl added to the nutrient solution of 5-day old seedlings for up to 24 h) and a 6-day salt treatment (seedlings grown in nutrient solution containing 200 m M NaCl for 6 days) were compared. Roots of intact seedlings were labeled in vivo with [³⁵S]-methionine. Poly(A)⁺ RNA was isolated and assayed in an in vitro translation system. The changes in the levels of the labeled polypeptides and translation products were analyzed following separation on two-dimensional polyacrylamide gels. Over the salt-shock time course, the majority of the changes in polypeptides and translation products were quantitative, as were the changes at 6 days. Qualitative changes occurred during the salt-shock time course, but were not observed at 6 days. The levels of polypeptides and translation products differed at each point of the salt-shock time course and at 6 days. In addition, changes were observed at the shortest time points examined, indicating a rapid response to salt.     

Light and electron microscope studies on pollen development in barley (Hordeum vulgare L.) grown under copper-sufficient and deficient conditions

Abstract. Pollen development in copper-deficient barley plants is highly irregular resulting in low and variable pollen fertility. The main cause of this sterility was found to be the abnormal development of the tapetum which becomes expansionary and invasive as the pollen develops. The ultrastructure of both tapetum and microspores is different from that of control material with irregularities of exine deposition, endopolyploidy of tapetal nuclei and an alteration of organelle composition being correlated with low fertility.     

Ultrastructural changes in aging leaves of a light-grown achlorophyllous mutant of barley

The pattern and sequence of cellular degradation during the course of leaf senescence remains obscure and the nature of the trigger that induces cell senescence is unknown. In order to probe the pre-mortem phase of senescence temporal changes in cell ultrastucture were studied in aging leaves of light-grown achlorophyllous Hordeum vulgare L. cv. Dyan mutant seedlings. Electron microscope examination of the ultrastructure of mesophyll cell plastids revealed the absence of ribosomes and a highly disorganized prolamellar body. Both the number and size of plastoglobuli increased with aging and this change coincided with depletion of starch grains and dilation of lamellar membranes. Aging of mesophyll cells occurred coincident with a decline in ribosome content of the cytoplasm and loss of matrix granularity. Loss of ribosomes associated with the outer nuclear envelope membrane and a reduction in chromatin were also apparent. Only after 10 days was there evidence of loss of internal membrane integrity and swelling of mitochondrial cristae. Compartmentation was thus maintained during the aging process with membrane dissolution occurring late in senescence. These results suggest that an inability to produce chlorophyll and carotenoids and form thylakoid stacks due to the absence of plastid ribosomes, contributes to the rapid onset of senescence in light-grown achlorophyllous seedlings. Furthermore, disruption of chloroplast ribosome synthesis/assembly may constitute part of the plastid signal involved in triggering cell senescence.     

Modulation of leaf elongation, tiller appearance and tiller senescence in spring barley by far-red light

Supplemental far-red (FR) illumination of light-grown grass seedlings inhibits tiller production while enhancing leaf elongation. Although much is known about FR enhancement of internode elongation in dicots, relatively little research has been conducted to determine the effects of FR on monocot development. In growth chamber experiments, fibre optics were used to direct supplemental FR to elongating leaf blades, main stem bases and mature leaf blades of light-grown barley (Hordeum vulgare L.) seedlings. Our objective was to identify specific sites of perception for FR enhancement of leaf elongation and inhibition of tiller production, and to assess potential FR effects on tiller senescence. Far-red illumination of elongating leaves or of the main stem base reduced the total number of tillers per plant, primarily by reducing secondary and tertiary tiller production, and enhanced leaf elongation. However, leaf elongation was less sensitive to stem base treatments than to illumination of the elongating blade. Increased leaf length resulted from increased leaf elongation rate, while the duration of leaf elongation was unaffected. Exposure of mature leaf blades to FR had no effect on tillering or leaf elongation. None of the FR treatments led to tiller senescence. Localization of FR perception in vertically oriented tissues such as elongating blades and stem bases permits early detection of reflected light from neighbouring plants, allowing rapid response to impending competition.     

Na-Ca interactions in barley seedlings: relationship to ion transport and growth

Abstract Salt-stressed plants often show Ca deficiency symptoms. The effects of NaCl salinity (1 to 150 mol m^-3) and supplemental Ca (10 mol m^-3) on Na and Ca transport in barley (Hordeum vulgare L.) and their relationship to growth were investigated. The adjustment of Na and Ca transport was investigated by examining young seedlings exposed to short-term (immediate) and long-term (7 d) exposure to salinity. When the plants were exposed to long-term treatments of salinity, the rate of sodium accumulation in roots was approximately 10 to 15% of short-term treatments. No significant adjustment in the transport to the shoot was observed. Rates of tracer (²²Na) transport were compared to calculated rates based on relative growth rates and tissue element concentrations. Comparisons between measured tracer and calculated rates of transport indicate that ²²Na transport may underestimate transport to the shoot because of dilution of the tracer in the root cytoplasm. Calcium uptake showed only minor adjustment with time. Measured rates of tracer transport to the shoot correlated well with calculated values. The transport and tissue concentrations of Na were significantly affected by supplemental Ca. Calcium transport and tissue concentrations were markedly inhibited by salinity. Supplemental Ca increased Ca transport and accumulation at all NaCl treatments above that of control plants without supplemental Ca. Salinity inhibited plant growth at 150 mol m ^-3NaCl, but not at 75 mol m^-3. Supplemental Ca significantly improved root length but not fresh weight after 7d of salinity, although differences in fresh weight were detected after 9d. There were significant Na-Ca interactions with ion transport, ion accumulation, and growth. The effects of salinity on Na and Ca transport to the shoot do not appear to play a major role in shoot growth of barley.     

Potassium channels in barley: cloning, functional characterization and expression analyses in relation to leaf growth and development

It is not known how the uptake and retention of the key osmolyte K⁺ in cells are mediated in growing leaf tissue. In the present study on the growing leaf 3 of barley, we have cloned the full-length coding sequence of three genes which encode putative K⁺ channels ( HvAKT1 , HvAKT2 , HvKCO1 / HvTPK1 ), and of one gene which encodes a putative K⁺ transporter ( HvHAK4 ). The functionality of the gene products of HvAKT1 and HvAKT2 was tested through expression in Xenopus laevis oocytes. Both are inward-rectifying K⁺ channels which are inhibited by Cs⁺. Function of HvAKT1 in oocytes requires co-expression of a calcineurin-interacting protein kinase ( At CIPK23) and a calcineurin B-like protein (AtCBL9) from Arabidopsis , showing cross-species complementation of function. In planta , HvAKT1 is expressed primarily in roots, but is also expressed in leaf tissue. HvAKT2 is expressed particularly in leaf tissue, and HvHAK4 is expressed particularly in growing leaf tissue. Within leaves, HvAKT1 and HvAKT2 are expressed predominantly in mesophyll. Expression of genes changes little in response to low external K⁺ or salinity, despite major changes in K⁺ concentrations and osmolality of cells. Possible contributions of HvAKT1 , HvAKT2 , HvKCO1 and HvHAK4 to regulation of K⁺ relations of growing barley leaf cells are discussed.     

Far-red light reflected from neighbouring vegetation promotes shoot elongation and accelerates flowering in spring barley plants

Field experiments were conducted in St Paul, MN, USA, to test the hypothesis that early season declines in the red:far-red ratio (R:FR) associated with FR reflection from neighbouring leaves have a role in regulating barley development. Treatment plants were grown adjacent to densely sown border rows of barley. The borders functioned to reflect far-red (FR), which reduced R:FR within the treatment plant light environment without shading treatment plants. Barriers were set in the soil to minimize root interactions between treatment plants and borders. Treatment plants were spaced either 2 or 16 cm apart. The presence of borders significantly increased shoot leaf and internode lengths at both plant spacings. Leaf sheath length data suggest that interactions between 2 cm spaced treatment plants enhanced plant responsiveness to the presence of borders. Border treatments shortened the period of vegetative growth prior to initiation of main shoot floral primordia. Bordered plants formed fewer main shoot leaves, initiated internode elongation at a lower node, and had slightly earlier heading dates than unbordered controls. Leaf appearance rate was not influenced by border treatments. We conclude that barley shoot development is photomorphogenically modulated by R:FR. Early season shifts in R:FR could have a significant influence on shoot development given that barley has the capacity to detect and developmentally respond to declines in R:FR associated with FR reflection from neighbours.     

Changes in water relations and endogenous abscisic acid content of wheat and barley grains and embryos during development

Abstract. Immature cereal embryo development can be controlled by in vitro culture on media containing ABA, or by media of low osmotic potential. To assess the possible in vivo roles of these factors, endogenous ABA levels and water relations of embryos and grains of wheat ( Triticum aestivum L.) and barley ( Hordeum vulgare L.) were determined during development. ABA concentrations remained consistent with those required to inhibit precocious germination in vitro of early stage embryos but not of more mature embryos. With increasing maturity, a difference in water potential developed between grain and embryo, suggestive of an in vivo role for water status in controlling the development of the embryo.     

XET-related genes and growth kinematics in barley leaves

DV, displacement velocity
GA₃, gibberellic acid
REGR, relative elemental growth rate
XET, xyloglucan-endotransglycosylase

Recently Schünmann et al. (1997 ; Plant, Cell and Environment 20, 1439–1450) investigated the correlation of spatial patterns of xyloglucan-endotransglycosylase (XET) activity, XET-related mRNAs, and growth in elongating barley ( Hordeum vulgare L.) leaves. Here, methodological difficulties in the kinematic growth analysis are discussed, and it is concluded that the role that XET-related gene activity plays in the control of spatial growth patterns remains undetermined.     

The base of the leaf acts as a localized sink for photosynthate in mature barley leaves

The gradients in photosynthetic and carbohydrate metabolism which persist within the fully expanded second leaf of barley ( Hordeum vulgare ) were examined. Although all regions of the leaf blade were green and photosynthetically active, the basal 5 cm, representing approximately 20% of the leaf area, retained some characteristics of sink tissue. The leaf blade distal from the leaf sheath exhibited characteristics typical of source tissue; the activities of sucrolytic enzymes (invertase and sucrose synthase) were relatively low, whilst that of sucrose phosphate synthase was high. These regions of the leaf accumulated sucrose throughout the photoperiod and starch only in the second half of the photoperiod whilst hexose sugars remained low. By contrast the leaf blade proximal to the leaf sheath retained relatively high activities of sucrolytic enzymes (especially soluble, acid invertase) whilst sucrose phosphate synthase activity was low. Glucose, as well as sucrose, accumulated throughout the photoperiod. Although starch accumulated in the second half of the photoperiod, a basal level of starch was present throughout the photoperiod, by contrast with the rest of the leaf. The ¹⁴CO₂ feeding experiments indicated that a constant amount of photosynthate was partitioned towards starch in this region of the leaf irrespective of irradiance. These findings are interpreted as the base of the leaf blade acting as a localized sink for carbohydrate as a result of sucrose hydrolysis by acid invertase.     

Accumulation of glycinebetaine during cold acclimation and freezing tolerance in leaves of winter and spring barley plants

A study was performed to examine whether or not betaine (glycinebetaine), a compatible solute, is accumulated in response to cold stress and is involved in mechanisms that protect plants from freezing injury. For this purpose, we used near-isogenic lines of barley, with each line differing only in a single gene for the spring type of growth habit; the various lines were produced by back-crosses to a recurrent cultivar of the winter type. The winter type of growth habit requires a low temperature for triggering of flower development (vernalization), whereas the spring type does not. Betaine was accumulated to five times the basal level over the course of 3 weeks at low temperature (5 °C) in the winter-type cultivar and in a spring-sh line having the sh gene for the spring-type growth habit, but the level was only doubled in the spring-Sh3 line, which carried the Sh₃ gene for the spring-type growth habit. Among near-isogenic lines of the same cultivar, the levels of betaine accumulated in leaves at low temperature were well correlated with the percentages (on a dry weight basis) of green leaves that survived freezing injury (-5 °C). This observation indicates the possibility, separate from the recognized role of betaine in the response to salinity and/or drought, that betaine accumulates in response to cold stress and that the accumulation of betaine during cold acclimation is associated to some extent with freezing tolerance in leaves of barley plants.     

The relationship between leaf growth and ABA accumulation in the grass leaf elongation zone

Detached barley (Hordeum vulgare L.) shoots, maintained at different air temperatures and VPDs, were fed ABA via the sub-crown internode in a leaf elongation assay. Analysis of variance of leaf elongation rate (LER) showed significant effects of temperature (T), fed [ABA] and the interaction T × [ABA]. However, the interaction became non-significant when LER was modelled against the [ABA] of the elongation zone, [EZ-ABA] When detached barley shoots were fed sap from droughted maize (Zea mays L.) plants, sap [ABA] could not explain the growth inhibitory activity. Measurement of [EZ-ABA] accounted for this ‘unexplained’ growth inhibition. The detached shoot experiments indicated that [EZ-ABA], and not xylem sap [ABA], was an appropriate explanatory variable to measure in droughted plants. However, ABA accumulation in the elongation zone could not explain a 35% growth reduction in intact droughted plants; thus we considered an interaction of water status and ABA. Using a coleoptile growth assay, we applied mild osmotic stresses (ψ=0 to ?0.06 MPa) and 10^?4 mol m^?3 ABA. Individually, these treatments did not inhibit growth. However, osmotic stress and ABA applied together significantly reduced growth. This interaction may be an important mechanism in explaining leaf growth inhibition of droughted plants.     

Accumulation and subcellular distribution of cations in relation to the growth of potassium-deficient barley

Abstract Growth of barley (Hordeum vulgare L., cv. Georgie) was insensitive to soil K content above about 150 mg kg^?1, but at lower levels it declined. The reduction in yield was greater in soils containing approximately 10 mg Na kg^?1 than in soils with about 90 mg kg^?1 of Na. Growth was unaffected by changes in shoot K concentration above 75 mol m^?3, but declined at lower concentrations, and the decrease was less in plants grown in soils with high Na. Growth responses were not simply related to tissue K concentrations because plants grown in soils with extra Na had higher yields but lower K concentrations. When soil Na was low, plants accumulated Ca as tissue K declined, but when Na was provided this ion was accumulated. Plant Mg concentrations were generally low but increased as K decreased. The Ca and Mg were osmotically active. There were highly significant inverse linear relationships between yield and either the Ca or Mg concentrations in the shoots. X-ray microanalysis was used to examine the compartmentation of cations in leaves from barley plants (cv. Clipper) grown in nutrient solutions with high and low K concentrations. In plants grown with 2.5 mol m^?3 K, this was the major cation in both the cytoplasm and vacuole of mesophyll cells. However, in plants grown with 0.02 mol m^?3 K it declined to undetectable levels in the vacuole, although it was still detectable in the cytoplasm. In all plants, Ca was mainly located in epidermal cells. The implication of the results for explaining responses to K. in terms of compartmentation of solutes is discussed.     

Photosynthesis and translocation rate in high lysine mutant barley

Photosynthesis and translocation rates were studied in high lysine barley mutant Notch-2 and its parent NP 113. Photosynthesis rates were higher in the     

Drought- and ABA-induced changes in photosynthesis of barley plants

The changes caused by drought stress and abscisic acid (ABA) on photosynthesis of barley plants (Hordeum vulgare. L. cv. Alfa) have been studied. Drought stress was induced by allowing the leaves to lose 12% of their fresh weight. Cycloheximide (CHI), an inhibitor of stress-induced ABA accumulation, was used to distinguish alterations in photosynthetic reactions that are induced after drought stress in response to elevated ABA levels from those that are caused directly by altered water relations. Four hoars after imposition of drought stress or 2 h after application of ABA, Ihe bulk of the leaf's ABA content measured by enzyme-amplified ELISA, increased 14- and 16-fold, respectively. CHI fully blocked the stress-induced ABA accumulation. Gas exchange measurements and analysis of enzyme activities were used to study the reactions of photosynthesis to drought stress and ABA. Leaf dehydration or ABA treatment led to a noticeable decrease in both the initial slope of the curves representing net photosynthetic rate versus intercellular CO₂ concentration and the maximal rate of photosynthesis; dehydration of CHI-treated plants showed much slower inhibition of the latter. The calculated values of the intercellular CO₂ concentration, CO₂ compensation point and maximal carboxylating efficiency of ribulose 1,5-bisphosphate (RuBP) carboxylase support the suggestion that biochemical factors are involved in the response of photosynthesis to ABA and drought stress. RuBP carboxylase activity was almost unaffected in ABA- and CHI-treated, non-stressed plants. A drop in enzyme activity was observed after leaf dehydration of the control and ABA-treated plants. When barley plants were supplied with ABA, the activity of carbonic anhydrase (CA, EC 4.2.2.1) increased more than 2-fold. Subsequent dehydration caused an over 1.5-fold increase in CA activity of the control plants and a more than 2.5-fold increase in ABA-treated plants. Dehydration of CHI-treated plants caused no change in enzyme activity. It is suggested that increased activity of CA is a photosynthetic response to elevated ABA concentration.     

Electrophysiological characterization of pathways for K+ uptake into growing and non-growing leaf cells of barley

Potassium is a major osmolyte used by plant cells. The accumulation rates of K⁺ in cells may limit the rate of expansion. In the present study, we investigated the involvement of ion channels in K⁺ uptake using patch clamp technique. Ion currents were quantified in protoplasts of the elongation and emerged blade zone of the developing leaf 3 of barley ( Hordeum vulgare L.). A time-dependent inward-rectifying K⁺-selective current was observed almost exclusively in elongation zone protoplasts. The current showed characteristics typical of Shaker-type channels. Instantaneous inward current was highest in the epidermis of the emerged blade and selective for Na⁺ over K⁺. Selectivity disappeared, and currents decreased or remained the same, depending on tissue, in response to salt treatment. Net accumulation rates of K⁺ in cells calculated from patch clamp current–voltage curves exceeded rates calculated from membrane potential and K⁺ concentrations of cells measured in planta by factor 2.5–2.7 at physiological apoplastic K⁺ concentrations (10–100 m m ). It is concluded that under these conditions, K⁺ accumulation in growing barley leaf cells is not limited by transport properties of cells. Under saline conditions, down-regulation of voltage-independent channels may reduce the capacity for growth-related K⁺ accumulation.     

Acclimatization of K+ uptake to changes in root temperature: experiments with oilseed rape and barley in flowing solution culture

Abstract Changes in the net uptake rate of K⁺ and in the average tissue concentration of K⁺ were measured over 14 d in response to changes in root temperature with oilseed rape (Brassica napus L. cv. Bien venu) and barley (Hordeum vulgare L. cv. Atem). Plants were grown in flowing nutrient solutions containing 2.5 mmol m^?3 K⁺ and were acclimatized over 49 d (rape) or 28 d (barley) to low root temperature (5°C) prior to steady–state treatments at root temperatures between 3 °C and 25 °C, with common air temperature. Uptake of K⁺ was monitored continuously over 14 d and nitrogen was supplied as NH₄⁺+ NO^?₃ or NH⁺₄ or NO^?₃. Unit absorption rates of K⁺ increased with time and with root temperature up to Day 4 or 5 following the change in root temperature. Thereafter they usually approached steady-state, with Q₁₀? 2.0 between 7 °C and 17°C, although rates became similar between 7 °C and 13°C. Uptake of K⁺ by rape plants was invariably greater under NO^?₃ nutrition compared with NH⁺₄. The percentage K⁺ in the plant dry matter increased with temperature from 2% at 3 °C to 4% at 25 °C in rape, but there was less effect of temperature on the average concentrations of K⁺ in the plant fresh weight or plant water content. Concentrations of K⁺ in the leaf water fraction of rape plants decreased with increasing root temperature, but in barley they increased with increasing root temperature. Concentrations of K⁺ in the root water fraction were relatively stable with respect to root temperature. The results are discussed in terms of compensatory changes in K⁺ uptake following a change in root temperature and the relationships between growth, shoot: root ratio and K⁺ composition of the plant.