Nitrate assimilatory properties of barley grown under long-term N limitation: Effects of local nitrate supply in split-root cultures

The effect of nitrate availability on characteristics of the nitrate assimilatory system was investigated in N-limited barley (Hordeum valgare L. cv. Golf), grown with the seminal root system split into initially equal-sized halves. The cultures were continuously supplied with nitrate-N at a relative addition rate (RA) of 0.09 day^?1, which resulted in relative growth rates (RG) that were ca 85% of those observed under surplus nitrate nutrition. The total N addition was divided between the subroots in ratios of 100:0, 80:20, 70:30, 60:40, and 50:50. For comparison, standard cultures were grown at RAs ranging from 0.03 to 0.18 day^?1. Initially, biomass and N partitioning to the subroots responded strongly and proportionally to the nitrate distribution ratio. After 12-14 days no further effect was observed. The V_max for net nitrate uptake and in vitro nitrate reductase (NR) activity were measured in acclimated plants, i.e., after > 14 days under a certain nitrate regime. In subroots fed from 20 to 100% of the total N addition, V_max for net nitrate uptake increased slightly, whereas NR activity was unaffected. Uptake and NR activities were insignificant in the 0%-subroot. Uneven nitrate supply to individual subroots had negligible effect on the whole-plant ability for nitrate uptake, and the relative V_max (unit N taken up per unit N in whole plant tissue and time) remained about 7-fold in excess of the demand set by growth. Balancing nitrate concentrations (the resulting external nitrate concentrations at a certain RA) generally ranged between 2 and 10 μM at growth-limiting RA, both when predicted from uptake kinetics and when actually measured. When comparing split root and standard cultures when acclimated, it appears that uptake and NR activities in roots respond more strongly to over-all nitrate availability than to nitrate availability to individual subroots.     

Root biomass fraction as a function of growth degree days in wheat

Root, underground and above-ground biomass were measured on various wheat cultivars from 1986 to 1988 in the south-east of France. The results are expressed as root: total (f _r) or underground: total (f _u) biomass fractions. Observed f _r and f _u values are in good agreement with previous results. f _r and f _u decrease steadily from emergence to maturity, with an exponential tendency. When using cumulative growth degree days since emergence relative to cumulative growth degree days until ear emergence () as time scale, f _r and f _u can be expressed as simple functions of % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGceaqabeaacaWGMb% addaWgaaqaaiaadkhaaeqaamaabmaabaGccqaH4oqCdaahaaWcbeqa% aiaacQcaaaaamiaawIcacaGLPaaakiabg2da9iaaicdacaGGUaGaaG% imaiaaiwdacqGHRaWkcaaIWaGaaiOlaiaaiwdacaaI4aGaamyzamaa% CaaaleqabaGaeyOeI0IaaGymaiaac6cacaaI0aGaaGioaiabeI7aXn% aaCaaameqabaGaaiOkaaaaaaaakeaacaWGMbaddaWgaaqaaiaadwha% aeqaamaabmaabaGccqaH4oqCdaahaaWcbeqaaiaacQcaaaaamiaawI% cacaGLPaaakiabg2da9iaaicdacaGGUaGaaGymaiaaikdacqGHRaWk% caaIWaGaaiOlaiaaiIdacaaI4aGaamyzamaaCaaaleqabaGaeyOeI0% IaaGOmaiaac6cacaaIYaGaaGioaiabeI7aXnaaCaaameqabaGaaiOk% aaaaaaaaaaa!610D!\[\begin{gathered} f_r \left( {\theta ^* } \right) = 0.05 + 0.58e^{ - 1.48\theta ^* } \hfill \\ f_u \left( {\theta ^* } \right) = 0.12 + 0.88e^{ - 2.28\theta ^* } \hfill \\ \end{gathered} \]The incremental root biomass partitioning coefficient, % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaeqySde2aaS% baaSqaaiaadkhaaeqaaOGaeyypa0JaaiikaiaadsgacaWGxbWaaSba% aSqaaiaadkhaaeqaaOGaai4laiaadsgacaWG0bGaaiykaiaac+caca% GGOaGaamizaiaadEfadaWgaaWcbaGaamiDaaqabaGccaGGVaGaamiz% aiaadshacaGGPaaaaa!4834!\[\alpha _r = (dW_r /dt)/(dW_t /dt)\], which describes the net increase in root biomass dW _r over time dt relative to the increase in total biomass (dW _r) over the same time period, has been derived from f and the relative growth rate. Its time course is accurately represented by% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaeqySdegdda% WgaaqaaiaadkhaaeqaamaabmaabaGccqaH4oqCdaahaaWcbeqaaiaa% cQcaaaaamiaawIcacaGLPaaakiabg2da9iabgkHiTiaaicdacaGGUa% GaaGymaiaaiwdacqGHRaWkcaaIWaGaaiOlaiaaiAdacaaIZaGaamyz% amaaCaaaleqabaGaeyOeI0IaaGimaiaac6cacaaI5aGaaGioaiabeI% 7aXnaaCaaameqabaGaaiOkaaaaaaaaaa!4D15!\[\alpha _r \left( {\theta ^* } \right) = - 0.15 + 0.63e^{ - 0.98\theta ^* } \]Under our experimental conditions, with no severe water stresses or nutrient deficiencies, and for our sampling frequency, around 2 weeks, the development scale , is the main factor governing the time courses of f _r, f _u and _r.     

Influence of plant nutrient concentration on growth rate: Use of a nutrient interruption technique to determine critical concentrations of N,P and K in young plants

A method is described for determining the way in which growth rate varies with plant nutrient concentration using a simple nutrient interruption technique incorporating only 2 treatments. The method involves measuring the changes in growth and nutrient composition of otherwise well-nourished plants after the supply of one particular nutrient has been withheld. Critical concentrations are estimated from the relationship between the growth rate (expressed as a fraction of that for control plants of the same size which remained well-nourished throughout) and the concentration of the growth-limiting nutrient in the plants as deficiency developed. Trials of the method using young lettuce plants showed that shoot growth rate was directly proportional to total N (nitrate plus organic N) concentration, and linearly or near-linearly related to K and P concentration over a wide range; the corresponding relationship for nitrate was strongly curvi-linear. Critical concentrations (corresponding to a 10% reduction in growth rate) determined from these results were similar to critical values calculated from models derived from field data, but were generally higher than published estimates of critical concentration (based on reductions in shoot weight) for plants of a similar size. Reasons for these discrepancies are discussed. Nitrate, phosphate or potassium concentrations in sap from individual leaf petioles were highly sensitive to changes in shoot growth rate as deficiency developed, with the slope of the relationships varying with leaf position, due to differences both in their initial concentration and in the rates at which they were utilized in individual leaves. Each nutrient was always depleted more quickly in younger leaves than in older ones, providing earlier evidence of deficiency for diagnostic purposes. Although the plants were capable of accumulating nitrate, phosphate and potassium well in excess of that needed for optimum dry matter production during periods of adequate supply, the rate of mobilization of these reserves was insufficient to prevent reductions in growth rate as the plants became deficient. This brings into question the validity of the conventional concept that luxury consumption provides a store of nutrients which are freely available for use in times of shortage. The implications of these results for the use of plant analysis for assessing plant nutrient status are discussed.     

Indole-3-acetic acid levels after phytochrome-mediated changes in the stem elongation rate of dark- and light-grownPisum seedlings

The effect of red (R) and far-red (FR) light on stem elongation and indole-3-acetic acid (IAA) levels was examined in dwarf and tall Pisum sativum L. seedlings. Red light reduced the extension-growth rate of etiolated seedlings by 70–90% after 3 h, and this inhibition was reversible by FR. Inhibition occurred throughout the growing zone. After 3 h of R, the level of extractable IAA in whole stem sections from the growing zone of etiolated plants either increased or showed no change. By contrast, extractable IAA from epidermal peels consistently decreased 3 h after R treatments. Decreases of 40% were observed for epidermal peels from the top 1 cm of tall plants receiving 3 h R. Brief R treatments resulted in smaller decreases in epidermal IAA levels and these decreases were not as great when FR followed R. In lightgrown plants, end-of-day FR stimulated growth during the following dark period in a photoreversible manner. The uppermost 1 cm of expanding third internodes was most responsive to the FR. Extractable IAA from epidermal peels from the upper 1 cm of third internodes increased by 30% or more 5 h after FR. When R followed the FR the increases were smaller. Levels of IAA in whole stem sections did not change and were twofold greater than in dark-grown plants. In both dark- and light-grown tall plants, IAA levels were lower in epidermal peels than in whole stem segments. These results provide evidence that IAA is compartmentalized at the tissue level within the growing stem and that phytochrome regulation of stem elongation rates may be partly based on modulating the level of IAA within the epidermis.Abbreviations IAA indole-3-acetic acid - R red light - FR farred light We thank Yu-Xian Zhu for helping to develop methods for IAA analysis, James Reid for supplying the genetic lines of Pisum and Richard Cyr for the use of microscopy equipment. This work was supported by NSF grant DCB-8801880 and by Hatch funds from the College of Agriculture and Life Sciences at Cornell University. The gas chromatograph-mass spectrometer was funded by NSF grant DMB-8505974 and funds from the College of Agriculture and Life Sciences at Cornell University. A preliminary report of some of these experiments has appeared in Plant Growth Substances, 1991 (Behringer et al. 1992 b).     

Rapid calcium exchange for protons and potassium in cell walls of Chara

Net fluxes of Ca²⁺, H⁺ and K⁺ were measured from intact Chara australis cells and from isolated cell walls, using ion-selective microelectrodes. In both systems, a stimulation in Ca²⁺ efflux (up to 100 nmol m^?2 s^?1, from an influx of ～40 nmol m^?2 s^?1) was detected as the H⁺ or K⁺ concentration was progressively increased in the bathing solution (pH 7.0 to 4.6 or K⁺ 0.2 to 10mol m^?3, respectively). A Ca²⁺ influx of similar size occurred following the reverse changes. These fluxes decayed exponentially with a time constant of about 10 min. The threshold pH for Ca²⁺ efflux (pH 5.2) is similar to a reported pH threshold for acid-induced wall extensibility in a closely related characean species. Application of NH₄⁺ to intact cells caused prolonged H⁺ efflux and also transient Ca²⁺ efflux. We attribute all these net Ca²⁺ fluxes to exchange in the wall with H⁺ or K⁺. A theoretical treatment of the cell wall ion exchanges, using the ‘weak acid Donnan Manning’ (WADM) model, is given and it agrees well with the data. The role of Ca²⁺ in the cell wall and the effect of Ca²⁺ exchanges on the measured fluxes of other ions, including bathing medium acidification by H⁺ efflux, are discussed.     

Isolation and sequence analysis of the genomic DNA fragment encoding an aspartic proteinase inhibitor homologue from potato (Solanum tuberosum L.)

A genomic DNA clone encoding an aspartic proteinase inhibitor of potato was isolated from a lambda EMBL3 phage library using the aspartic proteinase inhibitor cDNA as a hybridization probe. The gene has all characteristic sequences normally found in eucaryotic genes. Typical CAAT and TATA box sequences were found in the 5-upstream region. In this part are also two putative regulatory AGGA box sequences located. In the genomic sequence there are no intron sequences interrupting the coding region. An open reading frame of the gene encodes a precursor protein of 217 amino acids which shows high percent identity with the aspartic proteinase inhibitor cDNA.     

Testing of different antibiotics against Gram-positive and Gram-negative bacteria isolated from plant tissue culture

Different Gram-positive and Gram-negative bacteria (Staphylococcus xylosus, S. aureus, S. cohnii, Bacillus sp., Corynebacterium sp., Pseudomonas vesicularis) were isolated from homogenized shoot tips of Drosera rotundifolia, Spatiphyllum sp., Syngonium cv. White butterfly, Nephrolepis exaltata cv. Teddy Junior. Growth inhibition of selected bacterial strains was examined using 28 different single antibiotics and 7 antibiotic mixtures. It was found that with the two mixtures Imipenem/Ampicillin and Imipenem/Penicillin G at concentrations of 5 mg l^–1 each, bacterial growth inhibition was most effective. Because of the lack of toxic effects on in vitro plants of 7 species it was proposed that these antibiotic mixtures can be applied advantageously to inhibit bacterial growth in tissue culture.