Effects of endophyte infection on drought stress tolerance of Lolium perenne accessions from the Mediterranean region

Perennial ryegrass (Lolium perenne) is a cool-season grass species that is often infected by the leaf-inhabiting endophyte Neotyphodium lolii. This particular endophyte is asexual and has the potential to impact host survival, growth and reproduction. The objective was to assess the potential costs or benefits of endophyte infection on drought stress tolerance of native perennial ryegrass accessions originally collected from Italy, Morocco, Tunisia, and Turkey. Sixty infected (E+) individuals from each accession were planted in a greenhouse. Half of these individuals were treated with a systemic fungicide to eliminate the endophyte (E−). For two drought periods water was withheld for 10-14 days and then allowed a one week recovery period following each. In some accessions under drought, E+ plants had more tillers, greater tiller lengths, total dry mass and green shoot mass than E− plants, suggesting a positive effect of endophyte infection on host growth. Total tiller length and the number of tillers showed significant population × treatment × infection interactions for 4 of 6 populations. This work is one of the few that documents the effects of endophyte infection for a common forage grass species from wild populations native to its distributional range. The results demonstrate that endophyte infection can help ameliorate abiotic stress such as drought and there may be a selective advantage for grasses from certain Mediterranean regions.     

Gas Exchange and Chlorophyll Fluorescence in Symbiotic and Non-Symbiotic Ryegrass Under Water Stress

The symbiotic association of endophyte fungus, Neotyphodium lolii, and ryegrass improves the ryegrass resistance to drought. This is shown by a 30 % increase in the number of suckers in infected plants (E+), compared to plants lacking endophyte (E−), and by a higher water potential in the E+ than E− plants. The E+ plants have higher stomatal conductance (g _s), transpiration rate, net photosynthetic rate (P _N), and photorespiratory electron transport rate than the E− plants. The maximal photochemical efficiency (F_v/F_m) and the actual photochemical efficiency (Φ_PS2) are not affected by the endophyte fungus. The increase in P _N of the E+ plants subjected to water stress was independent from internal CO₂ concentration. An increased P _N was observed in E+ plants also in optimal water supply. Hence the drought resistance of E+ plants results in increased g _s, P _N, and photorespiratory electron transport rate. This revised version was published online in June 2006 with corrections to the Cover Date.     

Neotyphodium lolii, a Fungal Leaf Endophyte, Reduces Fertility ofLolium perenneExposed to Elevated UV-B Radiation

Plants ofLolium perenne, grown with and without the balansoidfungal leaf endophyteNeotyphodium lolii, were exposed to threeultraviolet radiation treatments at an outdoor facility in theUK for 172 d. Plants were exposed to either (a) a 30% elevationabove the ambient erythemally-weighted level of UV-B (280–315nm) radiation under banks of cellulose diacetate filtered fluorescentlamps that also produce UV-A (315–400 nm) radiation (UV-B+A);(b) elevated UV-A radiation alone under banks of polyester filteredlamps; or (c) ambient levels of solar radiation under banksof unenergized lamps. The fertility of plants grown withN. loliiwassignificantly reduced by the elevated UV-B+A exposure. After172 d, these plants produced 70% fewer spikes, 75% fewer seeds,71% lower total weight of seed and 78% fewer seeds per g d.wt of plant tissue than plants colonized byN. loliiwhich wereexposed to ambient radiation. There was no discernible effectof elevated UV-B+A exposure on the fertility of endophyte-freeplants. Plants irradiated with UV-B+A developed 14% thickerleaves than those exposed to ambient radiation. Those whichwere irradiated with elevated UV-A alone produced seeds thatwere 20% heavier than plants exposed to ambient levels of radiation.Plants grown withN. loliihad 7% thicker leaves, 4% thicker stembases and 7% fewer tillers than those grown without it. Thefresh mass of tillers of plants grown withN. loliiwas 11% greaterthan those of endophyte-free plants, owing to their higher moisturecontents. These results suggest that the fertility ofL. perennecolonizedbyN. loliiin the natural environment could be deleteriouslyaffected by elevated fluxes of UV-B radiation associated withstratospheric ozone depletion and that this may affect the populationdynamics of the species.Copyright 1998 Annals of Botany Company Fungal leaf endophyte,Neotyphodium lolii, perennial ryegrass (Lolium perenne), stratospheric ozone depletion, UV-B radiation.     

Transformation of the ryegrass endophyte Neotyphodium lolii can alter its in planta mycelial morphology

The fungus Neotyphodium lolii grows in the intercellular spaces of perennial ryegrass as a mutualistic endosymbiont. One of the benefits it conveys to the plant is the production of alkaloids toxic to herbivores. We wanted to determine in planta expression patterns of the N. lolii 3-hydroxy-3-methylglutaryl-CoA reductase (HMG CoA reductase) gene, believed to be involved in the synthesis of two of these alkaloid toxins, lolitrem B and ergovaline. We transformed the N. lolii strain Lp19 with plasmids, in which DNA fragments upstream of the open reading frame of the N. lolii HMG CoA reductase gene controlled expression of the GUS (gusA; Escherichia coli β-glucuronidase) reporter gene. In exponentially growing cultures, the GUS gene was not expressed if the length of upstream sequence was less than 400 bp, and >1100 bp were required for maximum expression. When reintroduced into ryegrass plants, transformants often showed highly increased hyphal branching compared to the wild-type parent strain, although in culture their growth kinetics and morphology were indistinguishable from that of the wild-type. Deterioration of hyphae and the hypha–plant interface occurred and in one transformant reduced tillering (formation of new plants, referred to in agronomy as tillers) and death of infected plants. We found no evidence that these abnormalities were caused by interference of the construct with the function of the native gene, as judged by analysis of the site of integration of the promoter-GUS cassette, expression of the native gene and lolitrem B and ergovaline levels in infected plants. However, there was some correlation between GUS expression and the degree of hyphal branching, suggesting that high levels of β-glucuronidase may disturb the symbiotic interaction. Levels of another alkaloid, peramine, were also not significantly affected by transformation. In previous studies increased in planta branching of the endophyte has been shown to be associated with a severe reduction of alkaloid production. Our results show that a plant–endophyte association in which increased branching occurs is still able to produce alkaloids.     

Production of phenolics and the emission of volatile organic compounds by perennial ryegrass (Lolium perenne L.)/Neotyphodium lolii association as a response to infection by Fusarium poae

Grasses very often form symbiotic associations with Neotyphodium/Epichloë endophytic fungi. These endophytes often allow the host grass to be protected from different pathogens. However, there is little known about the mechanisms of such endophyte influence on the host. Thus, the purpose of this research was to examine the effect of the N. lolii endophyte on the total production of phenolic compounds, VOCs emission and the resistance of three perennial ryegrass genotypes infected by pathogenic Fusarium poae. Analyses of total phenolics content were performed in control (not inoculated) and inoculated plants after 1, 2, 3, 4, 5, and 6 days (DAI) and for VOCs after 0, 3, 6 and 12 DAI. The presence of endophytes significantly reduced the disease index in two of the three genotypes relative to that in E−. Plants infected by N. lolii exhibited higher production of phenolics relative to the E− plants. The highest amounts of phenolics were observed on the second and sixth DAI. Genotype Nl22 showed the strongest effect of the endophyte on the production of phenolics, which increased by over 61%. Both the endophyte infected and non-infected plants emitted most abundantly two GLVs ((Z)-3-hexenal, (Z)-3-hexen-1-yl acetate), three terpenes (linalool, (Z)-ocimene, β-caryophyllene) and three shikimic acid pathway derivatives (benzyl acetate, indole, and methyl salicylate). The endophyte presence and the intervals of VOCs detection were a highly significant source of variation for all emitted volatiles (P < 0.001). The genotype of the perennial ryegrass significantly affected only the emission of methyl salicylate (P < 0.05) and β-caryophyllene (P < 0.05). Most of the VOCs ((Z)-3-hexen-1-yl acetate, (Z)-3-hexenal, linalool and methyl salicylate) reached their highest levels of emission on the sixth DAI, when averaged over genotypes and endophyte status. The results highlight the role of Neotyphodium spp. in the mediation of quadro-trophic interactions among plants, symbiotic endophytes, invertebrate herbivores and plant pathogenic fungi. Our results also confirm the fact that symbiotic plants can activate a defense reaction faster than non-symbiotic plants after a pathogen attack. Thus, N. lolii can be involved in the defense of perennial ryegrass against pathogens and potentially could be central to the host plants’ protection.     

Linear Relations between Carbon Dioxide Concentration and Rate of Development Towards Flowering in Sorghum, Cowpea and Soyabean

Negative linear relations were detected (P < 0·005)between the rate of progress from sowing to panicle initiationand CO₂ concentration (210-720 µmol CO₂ mol^-1 air) fortwo genotypes of sorghum [Sorghum bicolor (L.) Moench]. Relationsbetween CO₂ concentration and the rate of progress from sowingto first flowering were also negative in soyabean [Glycine max(L.) Merrill] (P < 0·025), but positive in cowpea[Vigna unguiculata (L.) Walp.] (P < 0·025), albeitthat in both grain legumes sensitivity was much less than insorghum. Thus CO₂ elevation does not delay flowering in allshort-day species. The considerable effect of CO₂ concentrationon times to panicle initiation resulted in large differencesamong the sorghum plants at this developmental stage; with increasein CO₂ concentration, plants were taller with slightly moreleaves and more pronounced apical extension. At the same timeafter sowing however, sorghum plants were heavier (P < 0·05)at 210 than at 360 µmol CO₂ mol^-1 air. In contrast, relationsbetween the dry masses of the soyabean and cowpea plants andCO₂ concentration were positive and curvilinear (P < 0·05).It is suggested that the impact of global environmental changecould be severe for sorghum production in the semi-arid tropics.Copyright1995, 1999 Academic Press Sorghum bicolor (L.) Moench., sorghum, Glycine max (L.) Merrill, soyabean, Vigna unguiculata (L.) Walp., cowpea, development, flowering, CO₂, dry matter accumulation, environmental change     

Effects of biotic and abiotic stress on the growth of three genotypes of Lolium perenne with and without infection by the fungal endophyte Neotyphodium lolii

The combinations of three genotypes of Lolium perenne with and without (i) infection by the fungal endophyte Neotyphodium lolii, (ii) infection by ryegrass mosaic virus and (iii) one of five different forms of abiotic stress were studied in pot experiments in a glasshouse. The five abiotic stress treatments were (i) low pH (compared with ‘optimal’ pH), (ii) cutting plants to a height of 1 cm (compared with 5 cm), (iii) shading (compared with no shading), (iv) cutting plants at 2‐weekly intervals (compared with 6 wk) and (v) low nitrogen applied (compared with ‘high’ nitrogen applied). On average, over the five experiments, the accumulated herbage dry weight was 10% more for N. lolii‐infected plants than uninfected, 22% more for virus‐free plants than infected, and 265% more for ‘unstressed’ plants than for plants with abiotic stress. The effects of N. lolii infection on plant growth when the plants were under abiotic or biotic stress were not consistent.     

Novel perennial ryegrass-Neotyphodium endophyte associations: relationships between seed weight,seedling vigour and endophyte presence

Two forage cultivars of perennial ryegrass (Lolium perenne) colonised by the mutualistic fungal endophyte Neotyphodium lolii, strain AR37, were used to investigate relationships between seed weight, seed vigour and endophyte presence. Seed was separated into six fractions according to weight, with each fraction divided into two further groups with the first being subject to accelerated ageing. Seed germination rates and proportions of viable and total endophyte frequencies were assessed for each fraction. Heavier fractions of seed produced a higher number of endophyte infected seedlings sooner than the lighter fractions for both cultivars. The highest proportion of viable endophyte was found in the fastest germinating perennial ryegrass seed for all weight fractions, from both cultivars, indicating a strong relationship between endophyte presence and seed germination rate. For one of the cultivars, after accelerated ageing, as seed weight increased the proportion of viable endophyte increased and the discrepancy between the proportion of endophyte in fresh and accelerated aged seed was reduced. This implies that for this cultivar heavier ryegrass seed provides a more favourable habitat to the dormant endophyte than lighter seed during storage and/or allowed for a greater biomass of endophyte hyphae to proliferate in the seed tissues prior to seed dormancy, thus allowing the fungus to develop more propagules and greater nutritional reserves.     

Genetic diversity and geographic differentiation in endangered Ammopiptanthus (Leguminosae) populations in desert regions of northwest China as revealed by ISSR analysis

• Background and Aims The desert legume genus Ammopiptanthuscomprises two currently endangered species, A. mongolicus andA. nanus. Genetic variability and genetic differentiation betweenthe two species and within each species were examined. • Methods Inter-simple sequence repeat (ISSR) marker datawere obtained and analysed with respect to genetic diversity,structure and gene flow. • Key Results Despite the morphological similarity betweenA. mongolicus and A. nanus, the two species are geneticallydistinct from each other, indicated by 63 % species-specificbands. Low genetic variability was detected for both populationlevel (Shannon indices of diversity H_pop = 0·106, percentageof polymorphic loci P = 18·55 % for A. mongolicus; H_pop= 0·070, P = 12·24 % for A. nanus) and specieslevel (H_sp = 0·1832, P = 39·39 % for A. mongolicus;H_sp = 0·1026, P = 25·89 % for A. nanus). Moderategenetic differentiation was found based on different measures(AMOVA _ST and Hickory ^B) in both A. mongolicus (0·3743–0·3744)and A. nanus (0·2162–0·2369). • Conclusions The significant genetic difference betweenthe two species might be due to a possible vicariant evolutionaryevent from a single common ancestor through the fragmentationof their common ancestor's range. Conservation strategies forthese two endangered species are proposed.     

Seasonal patterns of growth, dehydrins and water-soluble carbohydrates in genotypes of Dactylis glomerata varying in summer dormancy

• Background and Aims Summer dormancy in perennial grasseshas been studied inadequately, despite its potential to enhanceplant survival and persistence in Mediterranean areas. The aimof the present work was to characterize summer dormancy anddehydration tolerance in two cultivars of Dactylis glomerata(dormant ‘Kasbah’, non-dormant ‘Oasis’)and their hybrid using physiological indicators associated withthese traits. • Methods Dehydration tolerance was assessed in a glasshouseexperiment, while seasonal metabolic changes which produce putativeprotectants for drought, such as carbohydrates and dehydrinsthat might be associated with summer dormancy, were analysedin the field. • Key Results The genotypes differed in their ability tosurvive increasing soil water deficit: lethal soil water potential(_s) was –3·4 MPa for ‘Kasbah’ (althoughnon-dormant), –1·3 MPa for ‘Oasis’,and –1·6 MPa for their hybrid. In contrast, lethalwater content of apices was similar for all genotypes (approx.0·45 g H₂O g d. wt^–1), and hence the greater survivalof ‘Kasbah’ can be ascribed to better drought avoidancerather than dehydration tolerance. In autumn-sown plants, ‘Kasbah’had greatest dormancy, the hybrid was intermediate and ‘Oasis’had none. The more dormant the genotype, the lower the metabolicactivity during summer, and the earlier the activity declinedin spring. Decreased monosaccharide content was an early indicatorof dormancy induction. Accumulation of dehydrins did not correlatewith stress tolerance, but dehydrin content was a function ofthe water status of the tissues, irrespective of the soil moisture.A protein of approx. 55 kDa occurred in leaf bases of the mostdormant cultivar even in winter. • Conclusions Drought avoidance and summer dormancy arecorrelated but can be independently expressed. These traitsare heritable, allowing selection in breeding programmes.     

Conspicuous epiphytic growth of an interspecific hybrid Neotyphodium sp. endophyte on distorted host inflorescences

Selected Neotyphodium sp. endophytes are now commonly used to enhance pasture persistence and livestock productivity, with seed of perennial ryegrass and tall fescue cultivars with these selected endophytes being commercially available. In a large population of perennial ryegrass plants infected with a Neotyphodium sp. endophyte that was being grown for seed production a small percentage of inflorescences were distorted and covered with a conspicuous white mycelial growth. Within individual plants only a small number of inflorescences were affected and the amount of distortion differed between affected inflorescences. This Neotyphodium sp. is an interspecific hybrid of Epichloë typhina and Neotyphodium. lolii and like nearly all other Neotyphodium spp is symptomless in host grasses. The fungus isolated from distorted inflorescences had colonies that were identical to those isolated from symptomless inflorescences and these were characteristic of this Neotyphodium sp. This is the first report of distorted inflorescences covered with epiphytic hyphal growth on host grasses infected with an interspecific hybrid Neotyphodium sp.     

Allocation of Organ Biomass in Perfect and Imperfect Flowers

Perianth M_P, gynoecium M_G, and androecium M_A dry-weight biomass(in g) of 39 species of perfect flowers was measured. Thesedata were pooled with published data from an additional 51 speciesand used to determine size-dependent variations in (M_G and M_A)in terms of the hypothesis that the quotient of M_G and M_A exceeds1·0 for out-breeding (xenogamous) species and less than1·0 for in-breeding (autogamous) species. Ordinary leastsquare regression of the pooled data (n = 90) showed M_G = 0·118M^0·916_P (r² = 0·884) and M_A = 0·186 M^0·975_P(r² = 0·865), indicating that the biomass of the gynoeciumproportionally decrease as floral size increases. The exponentsof these regressions indicate that the ratio of gynoecial toandroecial biomass decreased with increasing floral size suchthat comparatively small flowers (M_P < 0·0021 g) hadM_G/M_A > 1·0 (predicted for 'out-breeders') while comparativelylarger flowers (M_P > 0·0021 g) had M_G /M_A < 1·0(predicted for 'in-breeders'). Thus, on average, the type ofbreeding system was a size-dependent phenomenon. To test whether the biomass of a floral organ-type is a legitimateindicator of gender reproductive effort, the biomass (in g)of stamen filaments M_m and anther sacs M_AS of 39 species wasdetermined. Least square regression of these data showed M_AS= 0·188 M^0·854_fil (r² = 0·967), indicatingthat species with larger stamen filaments, on the average, boreproportionally smaller anther sacs and thereby cautioning againstthe uncritical use of the allocation of biomass to floral organ-typeas a strict gauge of gender-function investment. To determine whether the loss of one gender-function resultsin proportional reallocation of biomass to the remaining gender-function,the size-dependency of androecial and gynoecial biomass wasdetermined for a total of 33 perfect and imperfect flowers ofCucumis melo. Regression of the data obtained from perfect flowersyielded M_A = 0·402 M^1·47_P (r² = 0·898)and M_G = 4·63 M^1·36_P (r² = 0·842). SinceM_G/M_A M^0·11_P , the biomass allocation to the gynoeciumrelative to the androecium decreased with increasing floralsize. This result was consistent with the broad interpecificcomparison based on 90 species with perfect flowers . Regressionof the data for imperfect flowers yielded M_A = 0·151M^1·02_P (r² = 0·675) and M_G = 4·68 M^1·47_P(r² = 0·996), indicating a near allometric relation forthe androecium and a strong positive anisometry for the gynoecium.Thus, for flowers of comparable size, a loss of female genderobtains a modest to significant again in androecial biomasswhereas the loss of male gender yields only a slight increasein gynoecial biomass. Collectively, the results of these studies indicate that biomassallocation patterns are size-dependent phenomena whose complexitieshave been largely ignored in the literature.Copyright 1993,1999 Academic Press Allometry, floral biomass, reproduction     

Relationship between Leaf Structure and Gas Exchange in Wheat Leaves at Different Insertion Levels

Net photosynthesis rate (P_n), stomatal conductance to CO₂ andresidual conductance to CO₂ were measured in the last six leaves(the sixth or flag leaf and the preceding five leaves) of Triticumaestivum L. cv. Kolibri plants grown in Mediterranean conditions.Recently fully expanded leaves of well-watered plants were alwaysused. Measurements were made at saturating photosynthetic photonflux density, and at ambient CO₂ and O₂ levels. The specificleaf area, total organic nitrogen content, some anatomical characteristics,and other parameters, were measured on the same leaves usedfor gas exchange experiments. A progressive xeromorphic adaptation in the leaf structure wasobserved with increasing leaf insertion levels. Furthermore,mesophyll cell volume per unit leaf area (V_mes/A) decreasedby 52·6% from the first leaf to the flag leaf. Mesophyllcell area per unit leaf area also decreased, but only by 24·5%.However, nitrogen content per unit mesophyll cell volume increasedby 50·6% from the first leaf to the flag leaf. This increasecould be associated to an observed higher number of chloroplastcross-sections per mm² of mesophyll cell cross-sectional areain the flag leaf: values of 23000 in the first leaf and 48000in the flag leaf were obtained. P_n per unit leaf area remainedfairly constant at the different insertion levels: values of33·83±0·93 mg dm^–2 h^–1 and32·32±1·61 mg dm^–2 h^–1 wereobtained for the first leaf and the flag leaf, respectively.Residual conductance, however, decreased by 18·2% fromthe first leaf to the flag leaf. Stomatal conductance increasedby 41·7%. The steadiness in P_n per unit leaf area across the leaf insertionlevels could be mainly accounted for by an opposing effect betweena decrease in V_mes/A and a more closely packed arrangement ofphotosynthetic apparatus. Adaptative significance of structuralchanges with increasing leaf insertion levels and the steadinessin P_n per unit leaf area was studied. Key words: Photosynthesis, structure, wheat     

Neotyphodium endophyte strain and superoxide dismutase activity in perennial ryegrass plants under water deficit

The presence of Neotyphodium endophyte in forage grass tillers has been associated with increased tolerance of abiotic stresses. The effect of four endophyte treatments (plants with three different strains of Neotyphodium lolii compared with plants without endophyte) on superoxide dismutase (SOD) (EC1.15.1.1) activity in Lolium perenne cv ‘Grasslands Samson’ was measured under high and low dehydration regimes in a glasshouse experiment. SOD activity was assayed by a microplate method utilising the inhibition of reduction of a tetrazolium dye by superoxide radicals. A progressive increase in dehydration over 2 weeks reduced shoot fresh weight, dry weight and shoot water content for high, compared with low, moisture-stressed plants. Mean shoot fresh weight was significantly lower for plants with strain AR37 endophyte than for plants with strain AR1 endophyte, wild-type or endophyte-free plants, but there was no interaction between endophyte treatment and dehydration treatment. There were no differences in mean SOD activity between the dehydration treatments, and the four endophyte treatments at any of the harvests. All harvest mean SOD levels for plants in both stress groups, however, were significantly different from the preceding harvest value. Between the first and second week of moisture stress there was a significant endophyte by harvest interaction for mean percentage change in SOD activity when activity in plants with wild-type strain endophyte increased more rapidly than in AR1, AR37 or endophyte-free plants. The results are in agreement with earlier reports suggesting that Neotyphodium endophytes do not have major effects on the water stress physiology of perennial ryegrass, although water deficits applied were not extreme.     

Modelling environmental variation in Young's modulus for Pinus radiata and implications for determination of critical buckling height

• Background and Aims Although density-specific stiffness,E/, (where E is Young's modulus and is wood density) is oftenassumed constant by the elastic similarity model, and in determinationof critical buckling height (H_crit), few studies have testedthis assumption within species. Here this assumption is testedfor Pinus radiata growing across an environmental gradient,and theory is combined with data to develop a model of Young'smodulus. • Methods Analyses use an extensive series of environmentalplots covering the range of climatic and edaphic conditionsover which P. radiata is grown in New Zealand. Reduced majoraxis regression was used to determine scaling exponents betweenlog–log plots of H_crit vs. groundline diameter (D), andE/ vs. D. Path analysis was used to identify significant directand indirect (through stem slenderness) edaphic and climaticinfluences on E. • Key Results Density-specific stiffness exhibited 3-foldvariation. As E/ scaled positively with D, the exponent of 0·95between H_crit and D exceeded the assumed value of 0·67under constant E/. The final path analysis model included meanair temperature in early autumn (T_aut) and slenderness as significant(P < 0·05) positive direct influences on E. Tree leafarea index and T_aut were indirectly associated with E throughtheir significant (P < 0·05) positive direct relationshipwith stem slenderness. Young's modulus was most sensitive toT_aut, followed by stem slenderness then leaf area index, andthe final model explained 76 % of the variance in E. • Conclusions The findings suggest that within speciesE/ variation may influence H_crit and the scaling exponent betweenD and H_crit so important in assumptions regarding allometricrelationships. The model presented may provide a useful meansof determining variation in E, E/ and H_crit across environmentalgradients.     

Nitrogen Utilization in N-limited Barley During Vegetative and Generative Growth: III. POST-ANTHESIS KINETICS OF NET NITRATE UPTAKE AND THE ROLE OF THE RELATIVE ROOT SIZE IN DETERMINING THE CAPACITY FOR NITRATE ACQUISITION

Barley (Hordeum vulgare L., cvs Golf, Mette, and Laevigatum)was grown under nitrogen limitation in solution culture untilnear maturity. Three different nitrogen addition regimes wereused: in the ‘HN’ culture the relative rate of nitrate-Naddition (RA) was 0·08 d^–1 until day 48 and thendecreased stepwise to, finally, 0·005 d^–1 duringgrain-filling; the ‘LN’ culture received 45% ofthe nitrogen added in HN; the ‘CN’ culture was maintainedat RA 0·0375 d^–1 throughout. Kinetics of net nitrateuptake were measured during ontogeny at 30 to 150 mmol m^–3external nitrate. V_max (which is argued to reflect the maximuminflux rate in these plants) declined with age in both HN andLN cultures. A pronounced transient drop was observed just beforeanthesis, which correlated in time with a peak in root nitrateconcentration. Similar, but less pronounced, trends were observedin CN. The relative V_max (unit nitrogen taken up per unit nitrogenin plants and day) in all three cultures declined from 1·3–2·3d^–1 during vegetative growth to 0·1–0·7d^–1 during generative growth. These values are in HN andLN cultures 15- to more than 100-fold in excess of the demandset by growth rates throughout ontogeny. Predicted balancingnitrate concentrations (defined as the nitrate concentrationrequired to support the observed rate of growth) were below6·0 mmol m^–3 in HN and LN cultures before anthesisand then decreased during ontogeny. In CN cultures the balancingnitrate concentration increased during grain-filling. Apartfrom the transient decline during anthesis, most of the effectof ageing on relative V_max can be explained in terms of reducedcontribution of roots to total biomass (R:T). The loss in uptakeper unit root weight is largely compensated for by the declinewith time in average tissue nitrogen concentrations. The quantitativerelationships between relative V_max and R:T in ageing plantsare similar to those observed for vegetative plants culturedat different RAs. The data support the contention that the capacity for nitrateacquisition in N-limited plants is under general growth control,rather than controlled by specific regulation of the biochemicalpathway of nitrate assimilation. Key words: Barley, nitrogen concentration, root: total plant biomass ratio, V_max     

Metabolomics analysis of the Lolium perenne–Neotyphodium lolii symbiosis: more than just alkaloids?

Above ground plant parts of Lolium perenne often harbour endophytic Neotyphodium lolii fungi. These occur both naturally and commercially, as variant strains are introduced to modify the grass metabolic profile. They reside in the apoplastic spaces and rarely cause visible symptoms of infection. The vast majority of literature has focussed on the biosynthesis, accumulation, and ecological relevance of a limited number of alkaloids produced by N. lolii which have been shown to negatively affect insect pests and vertebrate herbivores. Much less is known about the effects of other metabolites in these interactions or the role of resource supply on metabolic profiles, nor critically on the metabolic consequences of differences in the amount (concentration) of endophyte present. Here, we provide a synthesis of some of our recently published studies on effects of resource supply (nitrogen, carbohydrates) on concentrations of endophytes and endophyte specific metabolites in the L. perenne–N. lolii association. We present results of both quantitative PCR and targeted metabolomics studies, using contrasting endophyte strains in two perennial ryegrass cultivars. We also present and discuss a hypothetical schematic representation of possible links between plant and fungal metabolic networks. A multiple regression analysis of numerical insect responses and metabolic profiles indicates that effects of endophyte infection on insect population sizes could be predicted by concentrations of a range of metabolites other than alkaloids and depended on insect species, fungal strain, and nitrogen supply.     

The Effects of Elevated Atmospheric Carbon Dioxide and Water Stress on Ligth Interception, Dry Matter Production and Yield in Stands of Groundnut (Arachis hypogaea L.

Stands of groundnut (Arachis hypogaea L.), a C₃ legume, weregrown in controlled-environment glasshouses at 28 °C (±5°C)under two levels of atmospheric CO₂ (350 ppmv or 700 ppmv) andtwo levels of soil moisture (irrigated weekly or no water from35 d after sowing). Elevated CO₂ increased the maximum rate of net photosynthesisby up to 40%, with an increase in conversion coefficient forintercepted radiation of 30% (from 1–66 to 2–16g MJ^–1) in well-irrigated conditions, and 94% (from 0–64to 1·24 g MJ^–1) on a drying soil profile. In plantswell supplied with water, elevated CO₂ increased dry matteraccumulation by 16% (from 13·79 to 16·03 t ^–1) and pod yield by 25% (from 2·7 to 3·4t ha^–1).However, the harvest index (total poddry weight/above-grounddry weight) was unaffected by CO₂ treatment. The beneficial effects of elevated CO₂ were enhanced under severewater stress, dry matter production increased by 112% (from4·13 to 8·87 t ha^–1) and a pod yield of1·34t ha^–1 was obtained in elevated CO₂, whereascomparable plotsat 350 ppmv CO₂ only yielded 0·22 t ha^-1.There was a corresponding decrease in harvest index from 0·15to 0·05. Following the withholding of irrigation, plants growing on astored soil water profile in elevated CO₂ could maintain significantlyless negative leaf water potentials (P<0·01) for theremainder of the season than comparable plants grown in ambientCO₂, allowing prolonged plant activity during drought. In plants which were well supplied with water, allocation ofdry matter between leaves, stems, roots, and pods was similarin both CO₂ treatments. On a drying soil profile, allocationin plants grown in 350 ppmv CO₂ changed in favour of root developmentfar earlier in the season than plants grown at 700 ppmv CO₂,indicating that severe waterstress was reached earlier at 350ppmv CO₂. The primary effects of elevated CO₂ on growth and yield of groundnutstands weremediated by an increase in the conversion coefficientfor intercepted radiation and the prolonged maintenance of higherleaf water potentials during increasing drought stress. Key words: Arachis hypogaea, elevated CO₂, water stress, dry matter production     

Nitrate-Ammonium Ratio Required for pH Homeostasis in Hydroponically Grown Soybean

Imsande, J. 1986. Nitrate-ammonium ratio required for pH homeostasisin hydroponically grown soybean.—J. exp. Bot. 37: 341–347. Plant acid-base homeostasis is achieved when the mmoles of hydroxylions produced in the plant equal the mmoles of protons. Reductionof nitrate to ammonia is the major source of hydroxyl ions whereasammonium uptake-assimilation and the metabolism of neutral sugarsto organic acids are the primary sources of protons. Soybean[Glycine max (L.) Merr plants were grown hydroponically on mediumsupplemented with 3·0 mol m^–3 nitrogen providedas various combinations of KNO₃ and NH₄NO₃ Plant growth consumedessentially all available nitrogen in each case; however, onlyin flasks supplemented with approximately 1·8 minolesof KNO₃ plus 0·6 mmole of NH₄NO₃ was the pH of the mediumunchanged. Thus, for every mmole of nitrogen assimilated, approximately0·6 mmole of dissociable protons must have been producedby the conversion of neutral sugars to carboxylic acids. Also,it was shown that a plant obtaining all of its nitrogen fromnitrate must neutralize or excrete approximately 0·5mmole of hydroxyl ion d^–1. Conversely, the plant derivingall of its nitrogen from dinitrogen must excrete or neutralizeat least 0·8 mmole of hydrogen ion d^–1 whereasthe plant deriving all of its nitrogen from aminonium must excreteor neutralize approximately 2·1 mmoles of hydrogen iond^–1. Nevertheless, plants grown on medium supplementedwith 2·4 mol m^–1 nitrate plus 0·6 mol m^–3ammonium did not achieve a higher growth rate than plants grownon 3·0 mol m^–3 nitrate. Key words: Glycine max, nitrogen fixation, nitrate utilization     

The Influence of NO-3 and NH+4 Nutrition on the Gas Exchange Characteristics of the Roots of Wheat (Triticum aestivum) and Maize (Zea mays) Plants

Respiratory oxygen consumption by roots was 1·4- and1·6-fold larger in NH⁺₄-fed than in NO^-₃-fed wheat (Triticumaestivum L.) and maize (Zea mays L.) plants respectively. Higherroot oxygen consumption in NH⁺₄-fed plants than in NO^-₃-fedplants was associated with higher total nitrogen contents inNH⁺4-fed plants. Root oxygen consumption was, however, not correlatedwith growth rates or shoot:root ratios. Carbon dioxide releasewas 1·4- and 1·2-fold larger in NO⁺3-fed thanin NH⁺₄-fed wheat and maize plants respectively. Differencesin oxygen and carbon dioxide gas exchange rates resulted inthe gas exchange quotients of NH^-₄-fed plants (wheat, 0·5;maize, 0·6) being greatly reduced compared with thoseof NO^-₃-fed plants (wheat, 1·0; maize, 1·1). Measuredrates of HCO^-₃ assimilation by PEPc in roots were considerablylarger in 4 mM NH⁺₄-fed than in 4 NO^-₃ plants (wheat, 2·6-fold;maize, 8·3-fold). These differences were, however, insufficientto account for the observed differences in root carbon dioxideflux and it is probable that HCO^-₃ uptake is also importantin determining carbon dioxide fluxes. Thus reduced root extension in NH⁺₄-fed compared with NO^-₃-fedwheat plants could not be ascribed to differences in carbondioxide losses from roots.Copyright 1993, 1999 Academic Press Triticum aestivum, wheat, Zea mays, maize assimilation, ammonium assimilation, root respiration