Quantitative distribution of gibberellins and indole-acetic acid in pea (Pisum sativum L.) seedlings

The objective was to determine the distribution in pea seedling of GA (by thin layer chromatography) and IAA (by HPLC). Three samples of 30 7/day-old seedlings were sectioned into: apical meristem + plumular hook; stem elongation zone; stem non-elongation zone; cotyledons; root non-elongation zone; root tip. The material was frozen in liquid N and stored at -80 degrees C. The stem elongation zone, the apical meristem+plumular hook and the root tip had low IAA, whereas the stem non-elongation zone and root non-elongation zone had high IAA content. The stem elongation zone and apical meristem plumular hook had high GA, while root non-elongation zone had the lowest GA content.

The effect of chilling on the photosynthetic activity in coffee (Coffea arabica L.) seedlings. The protective action of chloroplastid pigments

Coffea arabica is considered to be sensitive to low temperatures, being affected throughout its entire life cycle. Injury caused by chilling (low temperatures above zero degree centigrade) is characterized primarily by inhibition of the photosynthetic process. The objective of this work was to evaluate the role of photosynthetic pigments in the tolerance of coffee (C. arabica L.) seedlings to chilling. The evaluation the photosynthetic activity was made by emission of Chl a fluorescence at room temperature (25°C) in vivo and in situ, using a portable fluorometer. The pigment content was obtained by extraction with 80% acetone, while estimation of membrane lipid peroxidation was determined by measuring the MDA content in leaf tissue extracts. The results indicated a generalized reduction in the quantum yield of PSII when the seedlings were maintained in the dark. The reduction occurred in the seedlings submitted to chilling treatment as well as in the control ones. This demonstrates that not only chilling acts to cause an alteration in PSII. It is possible that the tissue storage reserves had been totally exhausted, with the respiratory rate exceeding the photosynthetic rate; the later was nil, since the seedlings were kept in the dark. The efficiency in the capture, transfer and utilization of light energy in PS11 photochemical reactions requires a sequence of photochemical, biochemical and biophysical events which depend on the structural integrity of the photosynthetic apparatus. However, this efficiency was found to be related to the protective action of chloroplastid pigments, rather than to the concentration of these pigments.

Purification and Regulatory Properties of Mung Bean (Vigna Radiata L.) Serine Hydroxymethyltransferase 1

Serine hydroxymethyltransferase, the first enzyme in the pathway for interconversion of C1 fragments, was purified to homogeneity for the first time from any plant source. The enzyme from 72-h mung bean (Vigna radiata L.) seedlings was isolated using Blue Sepharose CL-6B and folate-AH-Sepharose-4B affinity matrices and had the highest specific activity (1.33 micromoles of HCHO formed per minute per milligram protein) reported hitherto. The enzyme preparation was extremely stable in the presence of folate or L-serine. Pyridoxal 5'-phosphate, ethylenediaminetetraacetate and 2-mercaptoethanol prevented the inactivation of the enzyme during purification. The enzyme functioned optimally at pH 8.5 and had two temperature maxima at 35 and 55°C. The Km values for serine were 1.25 and 68 millimolar, corresponding to Vmax values of 1.8 and 5.4 micromoles of HCHO formed per minute per milligram protein, respectively. The K0.5 value for L-tetrahydrofolate (H4folate) was 0.98 millimolar. Glycine, the product of the reaction and D-cycloserine, a structural analog of D-alanine, were linear competitive inhibitors with respect to L-serine with Ki values of 2.30 and 2.02 millimolar, respectively. Dichloromethotrexate, a substrate analog of H4folate was a competitive inhibitor when H4folate was the varied substrate. Results presented in this paper suggested that pyridoxal 5'-phosphate may not be essential for catalysis.The sigmoid saturation pattern of H4folate (nH = 2.0), one of the substrates, the abolition of sigmoidicity by NADH, an allosteric positive effector (nH = 1.0) and the increase in sigmoidicity by NAD+ and adenine nucleotides, negative allosteric effectors (nH = 2.4) clearly established that this key enzyme in the folate metabolism was an allosteric protein. Further support for this conclusion were the observations that (a) serine saturation exhibited an intermediary plateau region; (b) partial inhibition by methotrexate, aminopterin, O-phosphoserine, DL-{alpha}-methylserine and DL-O-methylserine; (c) subunit nature of the enzyme; and (d) decrease in the nH value from 2.0 for H4folate to 1.5 in presence of L-serine. These results highlight the regulatory nature of mung bean serine hydroxymethyltransferase and its possible involvement in the modulation of the interconversion of folate coenzymes.

Immunolocalization of glyoxysomal malate synthase from soybean cotyledons (Glycine max. L.)

Malate synthase (MS; EC 4.1.3.2), an enzyme specific to the glyoxylate cycle, was studied in cotyledons of dark-grown soybean (Glycine max L) seedlings with light and electron microscopy techniques. Immunogold localization confirmed biochemical evidence that MS from soybean is a glyoxysomal matrix enzyme.

Effect of the density and of the distance of Brachiaria decumbens staff on the initial growth of Coffea arabica L. Seedligns

The aim of this work was to evaluate the interference of three densities (4, 8 and 16 plants/m 2) of Brachiaria decumbens Stapf distributed at three different distances (0, 10 and 20 cm from coffee plants) on the initial growth of Coffea arabica L. seedlings. Dark red Latosoil was used as substrate to fill up 70 liters cement boxes. Coffee seedlings at 15 cm height and 8 leaves stage were planted in each box. B. decumbens seedlings were transplanted when they reached two to four leaves stage. The experimental design was a randomized blocks with ten treatments and three replications. At the end of the experiment were measured, height, leaf number, leaf area, chlorophyll content and dry weight of coffee plants. Dry weight of B. decumbens leaves was also evaluated. The leaf area and leaf dry weight was shown to be more sensitive parameters to the interference of Brachiaria plants. Dry weight of coffee plants were reduced from 60% to 88% when the B. decumbens were transplanted 0 cm from the coffee plant. With distance of 10 cm the reduction of dry weight increased with the increase of the density of the B. decumbens. In the coexistence of the coffee plants with 16 plants/m 2 of B. decumbens, there was a reduction in the dry weight of about 80% at the distances of 0 and 20 cm. Regarding leaf area, the reduction was 47% on the average in the density of 4 plants/m 2 of B. decumbens The reduction in leaf area at other densities was higher at the distance of 0 and 20 cm, and 55% reduction was obtained at the distance of 10 cm.

Removal of a combination of endocrine disruptors from aqueous systems by seedlings of radish and ryegrass

Endocrine disruptors (EDs) are widespread in the environment, especially aquatic systems, and cause dangerous effects on wildlife and humans. This work was aimed to assess the capacity of radish (Raphanus sativus L.) and ryegrass (Lolium perenne L.) seedlings to tolerate and remove two combinations of EDs containing bisphenol A (BPA), 17α-ethynilestradiol (EE2), and linuron from four aqueous media: distilled water, a solution of natural organic matter (NOM), a lake water and a river water. Seeds of the two species were germinated in each contaminated medium and, at the end of germination, the seedling growth was evaluated by biometric measurements and residual EDs were quantified by chromatographic analysis. Biometric measurements revealed that the phytotoxicity of the two combinations of EDs depended on the medium used. Radish showed a discrete tolerance in distilled water and lake water but was inhibited in the solution of NOM and river water. Ryegrass was negatively affected mainly in river water. The concentration of each ED appeared significantly reduced in all media in the presence of seedlings of both species, but not in the blanks without plants. In 5 days, radish removed up to 88% of BPA, 100% of EE2 and 42% of linuron, and in 6 days ryegrass removed up to 92% of BPA, 74% of EE2 and 16% of linuron. The considerable removal capacity of radish and ryegrass in all media tested encourages the use of phytoremediation to remove EDs from waters. © 2013 Copyright Taylor and Francis Group, LLC.

Changes in Salicylic Acid and Antioxidants during Induced Thermotolerance in Mustard Seedlings

Heat-acclimation or salicylic acid (SA) treatments were previously shown to induce thermotolerance in mustard (Sinapis alba L.) seedlings from 1.5 to 4 h after treatment. In the present study we investigated changes in endogenous SA and antioxidants in relation to induced thermotolerance. Thirty minutes into a 1-h heat-acclimation treatment glucosylated SA had increased 5.5-fold and then declined during the next 6 h. Increases in free SA were smaller (2-fold) but significant. Changes in antioxidants showed the following similarities after either heat-acclimation or SA treatment. The reduced-to-oxidized ascorbate ratio was 5-fold lower than the controls 1 h after treatment but recovered by 2 h. The glutathione pool became slightly more oxidized from 2 h after treatment. Glutathione reductase activity was more than 50% higher during the first 2 h. Activities of dehydroascorbate reductase and monodehydroascorbate reductase decreased by at least 25% during the first 2 h but were 20% to 60% higher than the control levels after 3 to 6 h. One hour after heat acclimation ascorbate peroxidase activity was increased by 30%. Young leaves appeared to be better protected by antioxidant enzymes following heat acclimation than the cotyledons or stem. Changes in endogenous SA and antioxidants may be involved in heat acclimation.

Anatomical and morphological responses of papaya, Carica papaya L., to various light conditions

Plants that develop under foliar shade encounter both low photosynthetically active radiation (PAR) and low red to far red ratios (R:FR). Both of these factors are important in determining developmental responses to shade. Papaya (Carica papaya L.) seedlings grown under filtered shade (low PAR and low R:FR) were compared with seedlings grown under neutral shade (low PAR with R:FR similar to that of full sunlight), and high light (moderate PAR with R:FR similar to that of full sunlight). The results indicated that papaya exhibits a light seeking strategy as evidenced by morphological and anatomical differences between treatments. Based on past research the results also indicate shade developmental responses in papaya to be phytochrome mediated.

Use of Sediment Bioassays to Verify Efficacy of Caulerpa taxifolia Eradication Treatments

Infestations of the marine macrophytic alga Caulerpa taxifolia were discovered in Agua Hedionda Lagoon, California in 2000. Rapid response actions included containment under pvc tarps coupled with injection of liquid sodium hypochlorite. To assess the efficacy of these treatments, replicated sediment cores were removed from representative treated sites and transferred to grow-out facilities. Similar cores from uninfested (control) sediments were removed, inoculated with viable explants of C. taxifolia and placed in grow-out facilities. Results from two sampling periods (1 year, 2 years post-treatment) showed that no viable C. taxifolia emerged in cores, and that inoculated “control” sediments supported normal growth. Eelgrass ( Zostera marina L.) seedlings emerged from native seed-banks in “treated” cores, which also supported growth of some invertebrates (annelid worms and hydroids). This study provided essential verification of C. taxifolia eradication efforts, and demonstrates the feasibility of incorporating quality control/quality assurance components in rapid response actions. Results of this study also suggest that seeds of eelgrass are viable for at least two years. (PDF has 9 pages.)

Ultrastructural alterations in mesophyll and bundle sheath chloroplasts of two maize cultivars in response to chilling at high irradiance

Maize (Zea mays L.) seedlings of two cultivars (cv. Bastion adapted to W. Europe, and cv. Batan 8686 adapted to the highlands of Mexico), raised in a glasshouse (19-25 degrees C), were transferred to 4.5 or 9 degrees C at photon flux density (PPFD) of 950 mu mol m(-2) s(-1) with 10-h photoperiod for 58 h and then allowed to recover at 22 degrees C for 16 h (14 h dark and 2 h at PPFD of 180 mu mol m(-2) s(-1)). The ultrastructural responses after 4 h or 26 h at 4.5 degrees C were the disappearance of starch grains in the bundle sheath chloroplasts and the contraction of intrathylakoid spaces in stromal thylakoids of the mesophyll chloroplasts. At this time, bundle sheath chloroplasts of cv. Batan 8686 formed peripheral reticulum. Prolonged stress at 4.5 degrees C (50 h) caused plastid swelling and the dilation of intrathylakoid spaces, mainly in mesophyll chloroplasts. Bundle sheath chloroplasts of cv. Batan 8686 seedlings appeared well preserved in shape and structure. Batan 8686 had also higher net photosynthetic rates during chilling and recovery than Bastion. Extended leaf photobleaching developed during the recovery period after chilling at 4.5 degrees C. This was associated with collapsed chloroplast envelopes, disintegrated chloroplasts and very poor staining.

Propagação vegetativa da alfavaca-cravo utilizando diferentes tipos de estacas e substratos

A propagação vegetativa é considerada importante ferramenta para o melhoramento de espécies lenhosas e herbáceas e vem sendo amplamente utilizada, visando melhorar e manter variedades de importância econômica e medicinal. Neste contexto utilizou-se estacas de alfavaca-cravo de 15 cm para apicais e 25 cm para as medianas, sendo estas com e sem folhas. As estacas foram plantadas em bandejas de isopor de 72 células, utilizando-se 10 substratos. Após 35 dias, as estacas foram retiradas com cuidado, sendo as partes aéreas e radiculares separadas e acondicionadas em saco de papel e mantidas em estufa a 70ºC por 48 h para posterior avaliação da matéria seca. Analisou-se a percentagem de enraizamento, número de folhas na estaca, peso de matéria seca de folha e de raiz. Não houve interação significativa entre os diferentes tipos de substrato e estaca para percentagem de enraizamento, sendo que os substratos de modo geral diferiram pouco entre si. Entre os tipos de estaca destacaram-se para esta variável a mediana sem folha e a apical com folha. Para as demais variáveis houve interação significativa entre o tipo de estaca e o substrato. A melhor estaca foi a mediana sem folhas plantadas em substrato arisco + esterco + húmus e vermiculita nas seguintes proporções: 20%:40%:40%; 40%:20%:40% e 40%:40%.