Biochemical dissection of diageotropica and Never ripe tomato mutants to Cd-stressful conditions

In order to further address the modulation of signaling pathways of stress responses and their relation to hormones, we used the ethylene-insensitive Never ripe (Nr) and the auxin-insensitive diageotropica (dgt) tomato mutants. The two mutants and the control Micro-Tom (MT) cultivar were grown over a 40-day period in the presence of Cd (0.2 mM CdCl2 and 1 mM CdCl2). Lipid peroxidation, leaf chlorophyll, praline content, Cd content and antioxidant enzyme activities in roots, leaves and fruits were determined. The overall results indicated that the MT genotype had the most pronounced Cd damage effects while Nr and dgt genotypes might withstand or avoid stress imposed by Cd. This fact may be attributed, at least in part, to the fact that the known auxin-stimulated ethylene production is comprised in dgt plants. Conversely, the Nr genotype was more affected by the Cd imposed stress than dgt, which may be explained by the fact that Nr retains a partial sensitivity to ethylene. These results add further information that should help unraveling the relative importance of ethylene in regulating the cell responses to stressful conditions. (C) 2012 Elsevier Masson SAS. All rights reserved.

Morphogenesis in guinea grass pastures under rotational grazing strategies

This study was conducted in order to evaluate the morphogenetic and structural characteristics of guinea grass cv. Mombasa under three post-grazing heights (intense - 30 cm, lenient - 50 cm and variable - 50 in spring-summer and 30 cm in autumn-winter) when sward light interception reached 95% during regrowth. Post-grazing heights were allocated to experimental units (0.25 ha) in a completely randomized block design with three replications. Post-grazing heights affected only leaf elongation rate and the number of live leaves. Pastures managed with variable post-grazing height showed higher leaf elongation rate in the summer of 2007. This management strategy also resulted in a higher number of live leaves. During the spring of 2006, plants showed lower leaf elongation rate, leaf appearance rate and number of live leaves, and greater phyllochron and leaf lifespan. In contrast, during the summer of 2007, the leaf appearance rate, leaf elongation rate, number of live leaves, and final leaf length were greater while phyllochron, stem elongation rate, and leaf senescence rate were lower. The management of the guinea grass cv. Mombasa with intense or variable post-grazing height throughout the year seems to represent an interesting management target, in terms of leaf appearance rate and number of live leaves.

Yield and chemical properties of rubber of Hevea clones according to phenological stages

The objective of this work was to evaluate the yield performance and macronutrient content of rubber extracted from four Hevea brasiliensis clones, under different tapping systems and plant phenological stages. The experiment was carried out in the 2010 and 2011 crop seasons, in a split-plot randomized complete block design, with four replicates. The main treatments - GT 1, PB 235, IAN 873, and RRIM 600 clones - were allocated in the plots, and the secondary treatments, which were the tapping systems 1/2S d/2, 1/2S d/4 ET 2.5%, and 1/2S d/7 ET 2.5%, were allocated in the subplots. The analyzed variables were natural rubber yield and macronutrient contents. Samples of natural rubber were obtained in the leaf development, mature leaf, and leaf senescence phenological stages. Rubber yield and its macronutrient contents are more influenced by tapping practice than by genetic material in the restrictive phenological stages of foliage.

Methodical study of nitrous oxide eddy covariance measurements using quantum cascade laser spectrometery over a Swiss forest

Nitrous oxide fluxes were measured at the Lägeren CarboEurope IP flux site over the multi-species mixed forest dominated by European beech and Norway spruce. Measurements were carried out during a four-week period in October–November 2005 during leaf senescence. Fluxes were measured with a standard ultrasonic anemometer in combination with a quantum cascade laser absorption spectrometer that measured N2O, CO2, and H2O mixing ratios simultaneously at 5 Hz time resolution. To distinguish insignificant fluxes from significant ones it is proposed to use a new approach based on the significance of the correlation coefficient between vertical wind speed and mixing ratio fluctuations. This procedure eliminated roughly 56% of our half-hourly fluxes. Based on the remaining, quality checked N2O fluxes we quantified the mean efflux at 0.8±0.4 μmol m−2 h−1 (mean ± standard error). Most of the contribution to the N2O flux occurred during a 6.5-h period starting 4.5 h before each precipitation event. No relation with precipitation amount could be found. Visibility data representing fog density and duration at the site indicate that wetting of the canopy may have as strong an effect on N2O effluxes as does below-ground microbial activity. It is speculated that above-ground N2O production from the senescing leaves at high moisture (fog, drizzle, onset of precipitation event) may be responsible for part of the measured flux.

Selection and hydroponic growth of bread wheat cultivars for bioregenerative life support systems

As part of the ESA-funded MELiSSA program, the suitability, the growth and the development of four bread wheat cultivars were investigated in hydroponic culture with the aim to incorporate such a cultivation system in an Environmental Control and Life Support System (ECLSS). Wheat plants can fulfill three major functions in space: (a) fixation of CO2 and production of O2, (b) production of grains for human nutrition and (c) production of cleaned water after condensation of the water vapor released from the plants by transpiration. Four spring wheat cultivars (Aletsch, Fiorina, Greina and CH Rubli) were grown hydroponically and compared with respect to growth and grain maturation properties. The height of the plants, the culture duration from germination to harvest, the quantity of water used, the number of fertile and non-fertile tillers as well as the quantity and quality of the grains harvested were considered. Mature grains could be harvested after around 160 days depending on the varieties. It became evident that the nutrient supply is crucial in this context and strongly affects leaf senescence and grain maturation. After a first experiment, the culture conditions were improved for the second experiment (stepwise decrease of EC after flowering, pH adjusted twice a week, less plants per m2) leading to a more favorable harvest (higher grain yield and harvest index). Considerably less green tillers without mature grains were present at harvest time in experiment 2 than in experiment 1. The harvest index for dry matter (including roots) ranged from 0.13 to 0.35 in experiment 1 and from 0.23 to 0.41 in experiment 2 with modified culture conditions. The thousand-grain weight for the four varieties ranged from 30.4 to 36.7 g in experiment 1 and from 33.2 to 39.1 g in experiment 2, while market samples were in the range of 39.4–46.9 g. Calcium levels in grains of the hydroponically grown wheat were similar to those from field-grown wheat, while potassium, magnesium, phosphorus, iron, zinc, copper, manganese and nickel levels tended to be higher in the grains of experimental plants. It remains a challenge for future experiments to further adapt the nutrient supply in order to improve senescence of vegetative plant parts, harvest index and the composition of bread wheat grains.

Extreme climatic events: impacts of drought and high temperature on physiological processes in agronomically important plants

Climate models predict more frequent and more severe extreme events (e.g., heat waves, extended drought periods, flooding) in many regions for the next decades. The impact of adverse environmental conditions on crop plants is ecologically and economically relevant. This review is focused on drought and heat effects on physiological status and productivity of agronomically important plants. Stomatal opening represents an important regulatory mechanism during drought and heat stress since it influences simultaneously water loss via transpiration and CO2 diffusion into the leaf apoplast which further is utilized in photosynthesis. Along with the reversible short-term control of stomatal opening, stomata and leaf epidermis may produce waxy deposits and irreversibly down-regulate the stomatal conductance and non-stomatal transpiration. As a consequence photosynthesis will be negatively affected. Rubisco activase—a key enzyme in keeping the Calvin cycle functional—is heat-sensitive and may become a limiting factor at elevated temperature. The accumulated reactive oxygen species (ROS) during stress represent an additional challenge under unfavorable conditions. Drought and heat cause accumulation of free amino acids which are partially converted into compatible solutes such as proline. This is accompanied by lower rates of both nitrate reduction and de novo amino acid biosynthesis. Protective proteins (e.g., dehydrins, chaperones, antioxidant enzymes or the key enzyme for proline biosynthesis) play an important role in leaves and may be present at higher levels under water deprivation or high temperatures. On the whole plant level, effects on long-distance translocation of solutes via xylem and phloem and on leaf senescence (e.g., anticipated, accelerated or delayed senescence) are important. The factors mentioned above are relevant for the overall performance of crops under drought and heat and must be considered for genotype selection and breeding programs.

Assessment of the ozone sensitivity of 22 native plant species from mediterranean annual pastures based on visible injury

Ozone (O3) phytototoxicity has been reported on a wide range of plantspecies, inducing the appearance of specific foliar injury or increasing leaf senescence. No information regarding the sensitivity of plantspecies from dehesa Mediterranean grasslands has been provided in spite of their great biological diversity. A screening study was carried out in open-top chambers (OTCs) to assess the O3-sensitivity of 22 representative therophytes of these ecosystems based on the appearance and extent of foliar injury. A distinction was made between specific O3injury and non-specific discolorations. Three O3 treatments (charcoal-filtered air, non-filtered air and non-filtered air supplemented with 40 nl l−1 O3 during 5 days per week) and three OTCs per treatment were used. The Papilionaceae species were more sensitive to O3 than the Poaceae species involved in the experiment since ambient levels induced foliar symptoms in 67% and 27%, respectively, of both plant families. An O3-sensitivity ranking of the species involved in the assessment is provided, which could be useful for bioindication programmes in Mediterranean areas. The assessed Trifoliumspecies were particularly sensitive since foliar symptoms were apparent in association with O3 accumulated exposures well below the current critical level for the prevention of this kind of effect. The exposure indices involving lower cut-off values (i.e. 30 nl l−1) were best related with the extent of O3-induced injury on these species.

Controlled Cytokinin Production in Transgenic Tobacco Using a Copper-Inducible Promoter

The cytokinin group of plant hormones regulates aspects of plant growth and development, including the release of lateral buds from apical dominance and the delay of senescence. In this work the native promoter of a cytokinin synthase gene (ipt) was removed and replaced with a Cu-controllable promoter. Tobacco (Nicotiana tabacum L. cv tabacum) transformed with this Cu-inducible ipt gene (Cu-ipt) was morphologically identical to controls under noninductive conditions in almost all lines produced. However, three lines grew in an altered state, which is indicative of cytokinin overproduction and was confirmed by a full cytokinin analysis of one of these lines. The in vitro treatment of morphologically normal Cu-ipt transformants with Cu2+ resulted in delayed leaf senescence and an increase in cytokinin concentration in the one line analyzed. In vivo, inductive conditions resulted in a significant release of lateral buds from apical dominance. The morphological changes seen during these experiments may reflect the spatial aspect of control exerted by this gene expression system, namely expression from the root tissue only. These results confirmed that endogenous cytokinin concentrations in tobacco transformants can be temporally and spatially controlled by the induction of ipt gene expression through the Cu-controllable gene-expression system.

Systemic gene expression in arabidopsis during an incompatible interaction with Alternaria brassicicola

Pathogen challenge can trigger an integrated set of signal transduction pathways, which ultimately leads to a state of high alert, otherwise known as systemic or induced resistance in tissue remote to the initial infection. Although large-scale gene expression during systemic acquired resistance, which is induced by salicylic acid or necrotizing pathogens has been previously reported using a bacterial pathogen, the nature of systemic defense responses triggered by an incompatible necrotrophic fungal pathogen is not known. We examined transcriptional changes that occur during systemic defense responses in Arabidopsis plants inoculated with the incompatible fungal pathogen Alternaria brassicicola. Substantial changes (2.00-fold and statistically significant) were demonstrated in distal tissue of inoculated plants for 35 genes (25 up-regulated and 10 down-regulated), and expression of a selected subset of systemically expressed genes was confirmed using real-time quantitative polymerase chain reaction. Genes with altered expression in distal tissue included those with putative functions in cellular housekeeping, indicating that plants modify these vital processes to facilitate a coordinated response to pathogen attack. Transcriptional up-regulation of genes encoding enzymes functioning in the beta-oxidation pathway of fatty acids was particularly interesting. Transcriptional up-regulation was also observed for genes involved in cell wall synthesis and modification and genes putatively involved in signal transduction. The results of this study, therefore, confirm the notion that distal tissue of a pathogen-challenged plant has a heightened preparedness for subsequent pathogen attacks.

Etr1-1 gene expression alters regeneration patterns in transgenic lettuce stimulating root formation

We have evaluated the transformation efficiency of two lettuce ( Lactuca sativa L.) cultivars, LE126 and Seagreen, using Agrobacterium tumefaciens- mediated gene transfer. Six- day- old cotyledons were co- cultivated with Agrobacterium cultures carrying binary vectors with two different genetic constructs. The first construct contained the beta- glucuronidase gene ( GUS) under the control of the cauliflower mosaic virus 35S promoter ( CaMV 35S), while the second construct contained the ethylene mutant receptor etr1- 1, which confers ethylene insensitivity, under the control of a leaf senescence- specific promoter ( sag12). Tissues co- cultivated with the GUS construct showed strong regeneration potential with over 90% of explants developing callus masses and 85% of the calli developing shoots. Histochemical GUS assays showed that 85.7% of the plants recovered were transgenic. Very different results were observed when cotyledon explants were co- cultivated with Agrobacteria carrying the etr1- 1 gene. There was a dramatic effect on the regeneration properties of the cultured explants with root formation taking place directly from the cotyledon tissue in 34% of the explants and no callus or shoots observed initially. Eventually callus formed in 10% of cotyledons and some organogenic shoots were obtained ( 2.86%). These results indicate that the ethylene insensitivity conferred by the etr1- 1 gene alters the normal pattern of regeneration in lettuce cotyledons, inhibiting the formation of shoots and stimulating root formation during regeneration.

Branching genes are conserved across species. Genes controlling a novel signal in pea are coregulated by other long-distance signals

Physiological and genetic studies with the ramosus (rms) mutants in garden pea (Pisum sativum) and more axillary shoots (max) mutants in Arabidopsis (Arabidopsis thaliana) have shown that shoot branching is regulated by a network of long-distance signals. Orthologous genes RMS1 and MAX4 control the synthesis of a novel graft-transmissible branching signal that may be a carotenoid derivative and acts as a branching inhibitor. In this study, we demonstrate further conservation of the branching control system by showing that MAX2 and MAX3 are orthologous to RMS4 and RMS5, respectively. This is consistent with the longstanding hypothesis that branching in pea is regulated by a novel long-distance signal produced by RMS1 and RMS5 and that RMS4 is implicated in the response to this signal. We examine RMS5 expression and show that it is more highly expressed relative to RMS1, but under similar transcriptional regulation as RMS1. Further expression studies support the hypothesis that RMS4 functions in shoot and rootstock and participates in the feedback regulation of RMS1 and RMS5 expression. This feedback involves a second novel long-distance signal that is lacking in rms2 mutants. RMS1 and RMS5 are also independently regulated by indole-3-acetic acid. RMS1, rather than RMS5, appears to be a key regulator of the branching inhibitor. This study presents new interactions between RMS genes and provides further evidence toward the ongoing elucidation of a model of axillary bud outgrowth in pea.

Unravelling the evolution of autumn colours: an interdisciplinary approach.

Leaf colour change is commonly observed in temperate deciduous forests in autumn. This is not simply a side effect of leaf senescence, and, in the past decade, several hypotheses have emerged to explain the evolution of autumn colours. Yet a lack of crosstalk between plant physiologists and evolutionary ecologists has resulted in slow progress, and so the adaptive value of this colour change remains a mystery. Here we provide an interdisciplinary summary of the current body of knowledge on autumn colours, and discuss unresolved issues and future avenues of research that might help reveal the evolutionary meaning of this spectacle of nature.

Desenvolvimento do capim-marandu com altura fixa ou variável durante as estações do ano

Aimed to identify, for assessing the tillering dynamics and morphogenesis , strategy (s ) suitable ( s) defoliation management to optimize the appearance of tillers and biomass of Urochloa Brizantha syn. Brachiaria brizantha cv. Marandu ( marandugrass ) throughout the year. The experiment was conducted from January 2013 to May 2014 were studied three strategies defoliation: Marandugrass with 30 cm throughout the experimental period (constant height); Marandugrass 15 cm in winter, the spring 30 cm and 45 cm in summer (increasing height); and Marandugrass 45 cm in winter, the spring 30 cm and 15 cm in summer (descending height). The experimental design was completely randomized in a split plot in time, with four replications. In winter, in the spring and summer, the following variables were calculated: appearance rate (TApP), mortality (TMoP) and survival (TSoP) of tillers, balance (BAL) between TApP and TMoP, stability index (IE) numbers of tillers and leaf elongation rates and stem, length of leaf blade and stem, leaf senescence rate, phyllochron and duration of life of the sheet. The TAPP was higher in the early spring and summer. For TMoP, the highest values were in the late spring and summer. The balance between TApP and TMoP was negative in winter and late spring, regardless of defoliation strategy. There was no difference in IE between the heights of the canopy during winter and late spring .. The number of tillers was higher in early spring and summer and lower in winter and late spring. The defoliation strategies have not changed phyllochron, leaf elongation rate. In spring and summer, there were increases in the rates of appearance, elongation and leaf senescence and, furthermore, decreased life span and phyllochron sheet. The plant height with increasing showed a lower rate of senescence and lower culm length in the spring. To increase the number of tillers marandu, it is advantageous to handle the plant with increasing height, ie, 15 cm in autumn and winter, the spring 30 cm and 45 cm in summer.The maintaining of B. brizantha cv. Marandu with fixed height of 30 cm or with variations between 15 and 45cm from the fall/winter to summer, does not influence the appearance and the growth of leaves per tiller, which indicates great flexibility on management defoliation in this forage plant.

Ecophysiology of Adonis distorta, a high-mountain species endemic of the Central Apennines

Morphological, anatomical and physiological plant and leaf traits of A. distorta, an endemic species of the Central Apennines on the Majella Massif, growing at 2,675 m a.s.l, were analyzed. The length of the phenological cycle starts immediately after the snowmelt at the end of May, lasting 128 ± 10 days. The low A. distorta height  (Hmax= 64 ± 4 mm) and total leaf area (TLA= 38 ± 9 cm2) associated to a high leaf mass area (LMA =11.8±0.6 mg cm−2) and a relatively high leaf tissue density (LTD = 124.6±14.3 mg cm−3) seem to be adaptive traits to the stress factors of the environment where it grows. From a physiological point of view, the high A. distorta photosynthetic rates (PN =19.6 ± 2.3 µmol m−2 s−1) and total chlorophyll content (Chla+b = 0.88 ± 0.13 mg g−1) in July are justified by the favorable temperature. PN decreases by 87% in September at the beginning of plant senescence. Photosynthesis and leaf respiration (RD) variations allow A. distorta to maintain a positive carbon balance during the growing season becoming indicative of the efficiency of plant carbon use. The results could be an important tool for conservation programmes of the A. distorta wild populations.

ANALYSIS OF POPULUS TREMULOIDES CLONAL VARIATION AND DELINEATION IN THE OTTAWA NATIONAL FOREST

The technique of delineating Populus tremuloides (Michx.) clonal colonies based on morphology and phenology has been utilized in many studies and forestry applications since the 1950s. Recently, the availability and robustness of molecular markers has challenged the validity of such approaches for accurate clonal identification. However, genetically sampling an entire stand is largely impractical or impossible. For that reason, it is often necessary to delineate putative genet boundaries for a more selective approach when genetically analyzing a clonal population. Here I re-evaluated the usefulness of phenotypic delineation by: (1) genetically identifying clonal colonies using nuclear microsatellite markers, (2) assessing phenotypic inter- and intraclonal agreement, and (3) determining the accuracy of visible characters to correctly assign ramets to their respective genets. The long-term soil productivity study plot 28 was chosen for analysis and is located in the Ottawa National Forest, MI (46° 37'60.0" N, 89° 12'42.7" W). In total, 32 genets were identified from 181 stems using seven microsatellite markers. The average genet size was 5.5 ramets and six of the largest were selected for phenotypic analyses. Phenotypic analyses included budbreak timing, DBH, bark thickness, bark color or brightness, leaf senescence, leaf serrations, and leaf length ratio. All phenotypic characters, except for DBH, were useful for the analysis of inter- and intraclonal variation and phenotypic delineation. Generally, phenotypic expression was related to genotype with multiple response permutation procedure (MRPP) intraclonal distance values ranging from 0.148 and 0.427 and an observed MRPP delta value=0.221 when the expected delta=0.5. The phenotypic traits, though, overlapped significantly among some clones. When stems were assigned into phenotypic groups, six phenotypic groups were identified with each group containing a dominant genotype or clonal colony. All phenotypic groups contained stems from at least two clonal colonies and no clonal colony was entirely contained within one phenotypic group. These results demonstrate that phenotype varies with genotype and stand clonality can be determined using phenotypic characters, but phenotypic delineation is less precise. I therefore recommend that some genetic identification follow any phenotypic delineation. The amount of genetic identification required for clonal confirmation is likely to vary based on stand and environmental conditions. Further analysis, however, is needed to test these findings in other forest stands and populations.