Effect of nitrogen supply on growth and photosynthesis of sunflower plants grown in the greenhouse

The effects of nitrogen availability on growth and photosynthesis were followed in plants of sunflower (Helianthus annuus L., var. CATISSOL-01) grown in the greenhouse under natural photoperiod. The sunflower plants were grown in vermiculite under two contrasting nitrogen supply, with nitrogen supplied as ammonium nitrate. Higher nitrogen concentration resulted in higher shoot dry matter production per plant and the effect was apparent from 29 days after sowing (DAS). The difference in dry matter production was mainly attributed to the effect of nitrogen on leaf production and on individual leaf dry matter. The specific leaf weight (SLW) was not affected by the nitrogen supply. The photosynthetic CO2 assimilation (A) of the target leaves was remarkably improved by high nitrogen nutrition. However, irrespective of nitrogen supply, the decline in photosynthetic CO2 assimilation occurred before the end of leaf growth. Although nitrogen did not change significantly stomatal conductance (gs), high-N grown plants had lower intercellular CO2 concentration (C-i) when compared with low-N grown plants. Transpiration rate (E) was increased in high-N grown plants only at the beginning of leaf growth. However, this not resulted in lower intrinsic water use efficiency (WUE). (C) 2004 Elsevier B.V.. All rights reserved.

Quantitative determination of anti-fungal and insecticide amides in adult plants, plantlets and callus from Piper tuberculatum by reverse-phase high-performance liquid chromatography

A rapid, sensitive and reliable reverse-phase HPLC method was used for the quantitative determination of the anti-fungal and insecticide amides, dihydropiplartine (1), piplartine (2), Delta(alpha,beta)-dihydropiperine (3) and pellitorine (4) in plants in natura, in plantlets in vitro and ex vitro, and in callus of Piper tuberculatum. Well-resolved peaks were obtained with good detection response and linearity in the range of 15.0-3000 mug/mL. The plants in natura contained compounds 1-4, the plantlets ex vitro and in vitro accumulated compounds 1-2 and 1-4, respectively, while only amide 4 was found in callus. Copyright (C) 2003 John Wiley Sons, Ltd.

Peroxidase enzyme (EC.1.11.1.7) activity as an indicator of water stress in sweet pepper plants

The present work was carried out at the Faculdade de Ciências Agronômicas - UNESP, Botucatu, SP. The purpose of the study was to evaluate the physiological and biochemical behavior of sweet pepper (Capsicum annuum L.) plants under different soil water availability conditions and the efficiency of the peroxidase (EC. 1.11.1.7) activity as an indicator of water stress in plants. Sweet pepper plants were grown for 230 days after transplanting of seedlings. The experiment was arranged in a completely randomized experimental design with 4 treatments, two irrigation managements (50 and 1500 kPa) and two soil surface managements (presence or absence of black polyethylene covering), and six replications. Physiological activities, such as stomatal transpiration and resistance to water vapor diffusion, were evaluated, as well as biochemical activities, such as peroxidase activity and total soluble protein in foliar tissues. It was observed that soil water availability may lead to physiological and biochemical alterations in plants. Successive water stress cycles may promote the development of characteristics responsible for improving the plant tolerance to periods of low water availability. The peroxidase enzyme activity showed to be an efficient indicator of water stress in sweet pepper plants.

Peroxidase activity as an indicator of water stress in sweet pepper plants

The purpose of the study was to evaluate the physiological and biochemical behavior of sweet pepper (Capsicum annuum L.) plants under different soil water availability conditions and the efficiency of the peroxidase (EC. 1.11. 1.7) activity as an indicator of water stress in plants. The experiment was carried out at the Faculdade de Ciências Agronômicas UNESP, Botucatu, SP. Sweet pepper plants were grown for 230 days after transplanting of seedlings and arranged in a completely randomized experimental design with 4 treatments, two irrigation managements (50 and 1500 kPa) and two soil surface managements (presence or absence of black polyethylene covering), and six replications. Physiological activities, such as stomatal transpiration and resistance to water vapor diffusion, were evaluated as well as biochemical activities, such as peroxidase activity and total soluble protein in foliar tissues. It was observed that soil water availability may lead to physiological and biochemical alterations in plants. Successive water stress cycles may promote the development of characteristics responsible for improving plant tolerance to periods of low water availability. The peroxidase enzyme activity showed to be an efficient indicator of water stress in sweet pepper plants.

Low abundance of long-tongued pollinators leads to pollen limitation in four specialized hawkmoth-pollinated plants in the Atlantic Rain forest, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

NUTRITIONAL STATUS AND LEVELS OF HEAVY METALS IN LETTUCE PLANTS FERTILIZED WITH ORGANIC COMPOUNDS

The use of organic compounds has been a good option to reduce spending on fertilizers, and gain increased productivity in the cultivation of lettuce. However, given the wide variety of raw materials used in the preparation of organic compounds, studies are needed to evaluate its effects on the release of essential nutrients to plants and on the release of contaminants such as heavy metals. The aim of this study was to evaluate the mineral nutrition and heavy metal contamination of lettuce in soils treated with doses and types of compost. The experiment was conducted in a greenhouse in randomized blocks in factorial scheme 5x4, with five types of organic compounds and four nitrogen levels (0, 35, 70 and 140 kg ha(-1) of N), with four replications. In general, the doses of the compounds were not enough to provide the necessary quantity of nutrients to the lettuce, with the exception of nitrogen. There was no increase in levels of heavy metals in the soil above that allowed by Brazilian legislation. Furthermore, compounds based on manure plus grass, and commercial compound caused increases in Zn concentration in plants at levels above the recommended for consumed.

PHOSPHITE AS PHOSPHORUS SOURCE TO GRAIN YIELD OF COMMON BEAN PLANTS GROWN IN SOILS UNDER LOW OR ADEQUATE PHOSPHATE AVAILABILITY

The effects of foliar and soil applied phosphite on grain yield in common bean (Phaseolus vulgaris L.) grown in a weathered soil under low and adequate phosphate availability were evaluated. In the first experiment, treatments were composed of a 2 x 7 + 2 factorial scheme, with 2 soil P levels supplied as phosphate (40 e 200 mg P dm(-3) soil), 7 soil P levels supplied as phosphite (0-100 mg P dm(-3) soil), and 2 additional treatments (without P supply in soil, and all P supplied as phosphite). In the second experiment, treatments were composed of a 2 x 3 x 2 factorial scheme, with 2 soil phosphate levels (40 e 200 mg P dm(-3) soil), combined with 3 nutrient sources applied via foliar sprays (potassium phosphite, potassium phosphate, and potassium chloride as a control), and 2 foliar application numbers (single and two application). Additional treatments showed that phosphite is not P source for common bean nutrition. Phosphite supply in soil increased the P content in shoot (at full physiological maturity stage) and grains, but at the same time considerably decreased grain yield, regardless of the soil phosphate availability. Foliar sprays of phosphite decreased grain yield in plants grown under low soil phosphate availability, but no effect was observed in plants grown under adequate soil phosphate availability. In general, foliar sprays of phosphate did not satisfactorily improve grain yield of the common bean plants grown under low soil phosphate availability.

Glyphosate has low toxicity to citrus plants growing in the field

Gravena, R., Filho, R. V., Alves, P. L. C. A., Mazzafera, P. and Gravena, A. R. 2012. Glyphosate has low toxicity to citrus plants growing in the field. Can. J. Plant Sci. 92: 119-127. There has been controversy over whether glyphosate used for weed management in citrus fields causes significant toxicity to citrus plants. Glyphosate may be toxic to non-target plants exposed to accidental application or drift. This work evaluated glyphosate toxicity in plants of Valencia citrus (Citrus sinensis. L. Osbeck) grafted onto 'Rangpur lime' (Citrus limonia L. Osbeck) and citrumelo 'Swingle' (Poncirus trifoliata (L.) Raf x Citrus paradisi Mad) by trunk- or foliar-directed herbicide applications under field conditions. In the first experiment, glyphosate was sprayed at rates of 0, 90, 180, 260, 540, 1080 and 2160 g a.e. ha(-1) directly on the trunk to a height of 5 cm above the grafting region. In the second experiment, glyphosate was sprayed on the plant canopies at rates of 0, 0.036, 0.36, 3.6, 36, 360 and 720 g a.e. ha(-1). There was no visual damage caused by glyphosate applied directly to the trunk, but the plants were affected by glyphosate sprayed directly on the canopies at rates over 360 g a.e. ha(-1). The main symptom was observed in the new shoots formed after the application, indicating an effect on meristems. Little or no effect was observed in mature leaves. Eight days after application the levels of shikimate, total free amino acids and total phenolic compounds were unaffected. All plants affected by glyphosate recovered between 6 and 12 mo after the treatments. Therefore, despite some transient symptoms Valencia citrus grafted onto 'Rangpur lime' and citrumelo 'Swingle' were tolerant to glyphosate.

Phosphite as phosphorus source to grain yield of common bean plants grown in soils under low or adequate phosphate availability

The effects of foliar and soil applied phosphite on grain yield in common bean (Phaseolus vulgaris L.) grown in a weathered soil under low and adequate phosphate availability were evaluated. In the first experiment, treatments were composed of a 2 x 7 + 2 factorial scheme, with 2 soil P levels supplied as phosphate (40 e 200 mg P dm-3 soil), 7 soil P levels supplied as phosphite (0-100 mg P dm-3 soil), and 2 additional treatments (without P supply in soil, and all P supplied as phosphite). In the second experiment, treatments were composed of a 2 x 3 x 2 factorial scheme, with 2 soil phosphate levels (40 e 200 mg P dm-3 soil), combined with 3 nutrient sources applied via foliar sprays (potassium phosphite, potassium phosphate, and potassium chloride as a control), and 2 foliar application numbers (single and two application). Additional treatments showed that phosphite is not P source for common bean nutrition. Phosphite supply in soil increased the P content in shoot (at full physiological maturity stage) and grains, but at the same time considerably decreased grain yield, regardless of the soil phosphate availability. Foliar sprays of phosphite decreased grain yield in plants grown under low soil phosphate availability, but no effect was observed in plants grown under adequate soil phosphate availability. In general, foliar sprays of phosphate did not satisfactorily improve grain yield of the common bean plants grown under low soil phosphate availability.

Genome-wide investigation reveals high evolutionary rates in annual model plants

Background: ;Rates of molecular evolution vary widely among species. While significant deviations from molecular clock have been found in many taxa, effects of life histories on molecular evolution are not fully understood. In plants, annual/perennial life history traits have long been suspected to influence the evolutionary rates at the molecular level. To date, however, the number of genes investigated on this subject is limited and the conclusions are mixed. To evaluate the possible heterogeneity in evolutionary rates between annual and perennial plants at the genomic level, we investigated 85 nuclear housekeeping genes, 10 non-housekeeping families, and 34 chloroplast;genes using the genomic data from model plants including Arabidopsis thaliana and Medicago truncatula for annuals and grape (Vitis vinifera) and popular (Populus trichocarpa) for perennials.;Results: ;According to the cross-comparisons among the four species, 74-82% of the nuclear genes and 71-97% of the chloroplast genes suggested higher rates of molecular evolution in the two annuals than those in the two perennials. The significant heterogeneity in evolutionary rate between annuals and perennials was consistently found both in nonsynonymous sites and synonymous sites. While a linear correlation of evolutionary rates in orthologous genes between species was observed in nonsynonymous sites, the correlation was weak or invisible in synonymous sites. This tendency was clearer in nuclear genes than in chloroplast genes, in which the overall;evolutionary rate was small. The slope of the regression line was consistently lower than unity, further confirming the higher evolutionary rate in annuals at the genomic level.;Conclusions: ;The higher evolutionary rate in annuals than in perennials appears to be a universal phenomenon both in nuclear and chloroplast genomes in the four dicot model plants we investigated. Therefore, such heterogeneity in evolutionary rate should result from factors that have genome-wide influence, most likely those associated with annual/perennial life history. Although we acknowledge current limitations of this kind of study, mainly due to a small sample size available and a distant taxonomic relationship of the model organisms, our results indicate that the genome-wide survey is a promising approach toward further understanding of the;mechanism determining the molecular evolutionary rate at the genomic level.

Sulfate assimilation in higher plants. Characterization of a stable intermediate in the adenosine 5′-phosphosulfate reductase reaction

The enzyme catalysing the reduction of adenosine 5′-phosphosulfate (AdoPS) to sulfite in higher plants, AdoPS reductase, is considered to be the key enzyme of assimilatory sulfate reduction. In order to address its reaction mechanism, the APR2 isoform of this enzyme from Arabidopsis thaliana was overexpressed in Escherichia coli and purified to homogeneity. Incubation of the enzyme with [35S]AdoPS at 4 °C resulted in radioactive labelling of the protein. Analysis of APR2 tryptic peptides revealed 35SO2–3 bound to Cys248, the only Cys conserved between AdoPS and prokaryotic phosphoadenosine 5′-phosphosulfate reductases. Consistent with this result, radioactivity could be released from the protein by incubation with thiols, inorganic sulfide and sulfite. The intermediate remained stable, however, after incubation with sulfate, oxidized glutathione or AdoPS. Because truncated APR2, missing the thioredoxin-like C-terminal part, could be labelled even at 37 °C, and because this intermediate was more stable than the complete protein, we conclude that the thioredoxin-like domain was required to release the bound SO2–3 from the intermediate. Taken together, these results demonstrate for the first time the binding of 35SO2–3 from [35S]AdoPS to AdoPS reductase and its subsequent release, and thus contribute to our understanding of the molecular mechanism of AdoPS reduction in plants.

Regulation of sulfur assimilation in higher plants: A sulfate transporter induced in sulfate-starved roots plays a central role in Arabidopsis thaliana

Proton/sulfate cotransporters in the plasma membranes are responsible for uptake of the environmental sulfate used in the sulfate assimilation pathway in plants. Here we report the cloning and characterization of an Arabidopsis thaliana gene, AST68, a new member of the sulfate transporter gene family in higher plants. Sequence analysis of cDNA and genomic clones of AST68 revealed that the AST68 gene is composed of 10 exons encoding a 677-aa polypeptide (74.1 kDa) that is able to functionally complement a Saccharomyces cerevisiae mutant lacking a sulfate transporter gene. Southern hybridization and restriction fragment length polymorphism mapping confirmed that AST68 is a single-copy gene that maps to the top arm of chromosome 5. Northern hybridization analysis of sulfate-starved plants indicated that the steady-state mRNA abundance of AST68 increased specifically in roots up to 9-fold by sulfate starvation. In situ hybridization experiments revealed that AST68 transcripts were accumulated in the central cylinder of sulfate-starved roots, but not in the xylem, endodermis, cortex, and epidermis. Among all the structural genes for sulfate assimilation, sulfate transporter (AST68), APS reductase (APR1), and serine acetyltransferase (SAT1) were inducible by sulfate starvation in A. thaliana. The sulfate transporter (AST68) exhibited the most intensive and specific response in roots, indicating that AST68 plays a central role in the regulation of sulfate assimilation in plants.

Overexpression of the Bacillus thuringiensis (Bt) Cry2Aa2 protein in chloroplasts confers resistance to plants against susceptible and Bt-resistant insects

Evolving levels of resistance in insects to the bioinsecticide Bacillus thuringiensis (Bt) can be dramatically reduced through the genetic engineering of chloroplasts in plants. When transgenic tobacco leaves expressing Cry2Aa2 protoxin in chloroplasts were fed to susceptible, Cry1A-resistant (20,000- to 40,000-fold) and Cry2Aa2-resistant (330- to 393-fold) tobacco budworm Heliothis virescens, cotton bollworm Helicoverpa zea, and the beet armyworm Spodoptera exigua, 100% mortality was observed against all insect species and strains. Cry2Aa2 was chosen for this study because of its toxicity to many economically important insect pests, relatively low levels of cross-resistance against Cry1A-resistant insects, and its expression as a protoxin instead of a toxin because of its relatively small size (65 kDa). Southern blot analysis confirmed stable integration of cry2Aa2 into all of the chloroplast genomes (5,000–10,000 copies per cell) of transgenic plants. Transformed tobacco leaves expressed Cry2Aa2 protoxin at levels between 2% and 3% of total soluble protein, 20- to 30-fold higher levels than current commercial nuclear transgenic plants. These results suggest that plants expressing high levels of a nonhomologous Bt protein should be able to overcome or at the very least, significantly delay, broad spectrum Bt-resistance development in the field.

A mammalian 2-5A system functions as an antiviral pathway in transgenic plants.

Resistance to virus infections in higher vertebrates is mediated in part through catalysis of RNA decay by the, interferon-regulated 2-5A system. A functional 2-5A system requires two enzymes, a 2-5A synthetase that produces 5'-phosphorylated, 2',5'-linked oligoadenylates (2-5A) in response to double-stranded RNA, and the 2-5A-dependent RNase L. We have coexpressed these human enzymes in transgenic tobacco plants by using a single plasmid containing the cDNAs for both human RNase L and a low molecular weight form of human 2-5A synthetase under control of different, constitutive promoters. Expression of the human cDNAs in the transgenic plants was demonstrated from Northern blots, by specific enzyme assays, and by immunodetection (for RNase L). Infection of leaves, detached or in planta, of the coexpressing transgenic plants by tobacco mosaic virus, alfalfa [correction of alfafa] mosaic virus, or tobacco etch virus resulted in necrotic lesions. In contrast, leaves expressing 2-5A synthetase or RNase L alone and leaves containing the plasmid vector alone produced typical systemic infections. While alfalfa mosaic virus produced lesions only in the inoculated leaves regardless of the concentration of virus in the inoculum, high, but not low, levels of tobacco etch virus inoculum resulted in escape of virus to uninoculated leaves. Nevertheless, there was a substantial reduction of tobacco etch virus yield as measured by ELISA assay in the coexpressing transgenic plants. These results indicate that expression of a mammalian 2-5A system in plants provides resistance to virus infections.

Mercuric ion reduction and resistance in transgenic Arabidopsis thaliana plants expressing a modified bacterial merA gene.

With global heavy metal contamination increasing, plants that can process heavy metals might provide efficient and ecologically sound approaches to sequestration and removal. Mercuric ion reductase, MerA, converts toxic Hg2+ to the less toxic, relatively inert metallic mercury (Hg0) The bacterial merA sequence is rich in CpG dinucleotides and has a highly skewed codon usage, both of which are particularly unfavorable to efficient expression in plants. We constructed a mutagenized merA sequence, merApe9, modifying the flanking region and 9% of the coding region and placing this sequence under control of plant regulatory elements. Transgenic Arabidopsis thaliana seeds expressing merApe9 germinated, and these seedlings grew, flowered, and set seed on medium containing HgCl2 concentrations of 25-100 microM (5-20 ppm), levels toxic to several controls. Transgenic merApe9 seedlings evolved considerable amounts of Hg0 relative to control plants. The rate of mercury evolution and the level of resistance were proportional to the steady-state mRNA level, confirming that resistance was due to expression of the MerApe9 enzyme. Plants and bacteria expressing merApe9 were also resistant to toxic levels of Au3+. These and other data suggest that there are potentially viable molecular genetic approaches to the phytoremediation of metal ion pollution.