The response of physiological and xylem anatomical traits under cadmium stress in Pinus thunbergii seedlings.

Studying the response of physiological and xylem anatomical traits under cadmium stress is helpful to understand plants' response to heavy metal stress. Here, seedlings of Pinus thunbergii Parl. were treated with 50, 100 and 150 mg kg-1 Cd2+ for 28 days. Cadmium and nonstructural carbohydrate content of leaves, stems and roots, root Cd2+ flux, cadmium distribution pattern in stem xylem and phloem, stem xylem hydraulic traits, cell wall component fractions of stems and roots, phytohormonal content such as abscisic acid, gibberellic acid 3, molecule -indole-3-acetic acid, and jasmonic acid from both leaves and roots, as well as xylem anatomical traits from both stems and roots were measured. Root Cd2+ flux increased from 50 to 100 mmol L-1 Cd2+ stress, however it decreased at 150 mmol L-1 Cd2+. Cellulose and hemicellulose in leaves, stems and roots did not change significantly under cadmium stress, while pectin decreased significantly. The nonstructural carbohydrate content of both leaves and stems showed significant changes under cadmium stress while the root nonstructural carbohydrate content was not affected. In both leaves and roots, the abscisic acid content significantly increased under cadmium stress, while the gibberellic acid 3, indole-3-acetic acid and jasmonic acid methylester content significantly decreased. Both xylem specific hydraulic conductivity and xylem water potential decreased with cadmium stress, however tracheid diameter and double wall thickness of the stems and roots were not affected. High cadmium intensity was found in both the stem xylem and phloem in all cadmium stressed treatments. Our study highlighted the in situ observation of cadmium distribution in both the xylem and phloem, and demonstrated the instant response of physiological traits such as xylem water potential, xylem specific hydraulic conductivity, root Cd2+ flux, nonstructural carbohydrate content, as well as phytohormonal content under cadmium stress, and the less affected traits such as xylem anatomical traits, cellulose and hemicellulose.

Correlation of biometric characteristics of fruits and seeds with the vigor of Agonandra brasiliensis seedlings in northern Amazonia

The goal of this study was to determine and correlate the main morphometric characteristics of fruits, seeds, and seedling vigor of Agonandra brasiliensis, in Roraima. The design was completely randomized with three treatments, consisting of six replications with 20 seeds. The treatments established were three classes of seeds: T1 - small seeds (S = 1.50-1.69 g); T2 - medium seeds (M = 1.70-1.89 g) and T3 - large seeds (L = 1.90-2.09 g). The variables evaluated were: length; diameter; the fresh mass of fruits and seeds; percentage, speed (in index) and average time of emergence, length of roots and aerial part; number of leaves; fresh, dry and total seedling mass; seedling lignification index; root/shoot ratio and dry root/shoot ratio and correlations between all studied variables. The fresh mass of the fruit of Agonandra brasiliensis has a high positive correlation with the mass of seeds. Small seeds of Agonandra brasiliensis exhibit a higher percentage of emergence in the conditions of the northern Amazon.

Calcium Phosphate Particles Coated with Humic Substances: A Potential Plant Biostimulant from Circular Economy.

Nowadays, the use of biostimulants to reduce agrochemical input is a major trend in agriculture. In this work, we report on calcium phosphate particles (CaP) recovered from the circular economy, combined with natural humic substances (HSs), to produce a plant biostimulant. CaPs were obtained by the thermal treatment of Salmo salar bones and were subsequently functionalized with HSs by soaking in a HS water solution. The obtained materials were characterized, showing that the functionalization with HS did not sort any effect on the bulk physicochemical properties of CaP, with the exception of the surface charge that was found to get more negative. Finally, the effect of the materials on nutrient uptake and translocation in the early stages of development (up to 20 days) of two model species of interest for horticulture, Valerianella locusta and Diplotaxis tenuifolia, was assessed. Both species exhibited a similar tendency to accumulate Ca and P in hypogeal tissues, but showed different reactions to the treatments in terms of translocation to the leaves. CaP and CaP-HS treatments lead to an increase of P accumulation in the leaves of D. tenuifolia, while the treatment with HS was found to increase only the concentration of Ca in V. locusta leaves. A low biostimulating effect on both plants' growth was observed, and was mainly scribed to the low concentration of HS in the tested materials. In the end, the obtained material showed promising results in virtue of its potential to elicit phosphorous uptake and foliar translocation by plants.

Transcriptomic and metabolomic profiling reveals the effect of LED light quality on morphological traits, and phenylpropanoid-derived compounds accumulation in Sarcandra glabra seedlings.

BACKGROUND: Sarcandra glabra is an evergreen and traditional Chinese herb with anti-oxidant, anti-bacterial, anti-inflammatory, and anti-tumor effects. Light is one of the most influential factor affecting the growth and quality of herbs. In recent times, the introduction of Light Emission Diode (LED) technology has been widely used for plants in greenhouse. However, the impact of such lights on plant growth and the regulatory mechanism of phenylpropanoid-derived compounds in S. glabra remain unclear. RESULTS: The red LED light (RL) substantially increased the plant height and decreased the stem diameter and leaf area relative to the white LED light (WL), while the blue LED light (BL) significantly reduced the height and leaf area of S. glabra. According to transcriptomic profiling, 861, 378, 47, 10,033, 7917, and 6379 differentially expressed genes (DEGs) were identified among the groups of leaf tissue under BL (BY) vs. leaf tissue under RL (RY), BY vs. leaf tissue under WL (WY), RY vs. WY, root tissue under WL (WG) vs. WY, stem tissue under WL (WJ) vs. WG, and WJ vs. WY, respectively. We identified 46 genes encoding for almost all known enzymes involved in phenylpropanoid biosynthesis, e.g., phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), and flavonol synthase (FLS). We found 53 genes encoding R2R3-MYB proteins and bHLH proteins, respectively, where several were related to flavonoids biosynthesis. A total of 454 metabolites were identified based on metabolomic profiling, of which 44, 87, and 296 compounds were differentially produced in WY vs. RY, WY vs. BY, and WY vs. WG. In BY there was a substantial reduction in the production of esculetin, caffeic acid, isofraxidin, and fraxidin, while the yields of quercitrin and kaempferol were significantly up-regulated. In RY, the contents of cryptochlorogenic acid, cinnamic acid, and kaempferol decreased significantly. Besides, in WG, the production of metabolites (e.g. chlorogenic acid, cryptochlorogenic acid, and scopolin) declined, while their yields increased significantly (e.g. esculetin, fraxetin, isofraxidin, and fraxidin). CONCLUSION: These results provide further insight into the regulatory mechanism of accumulation patterns of phenylpropanoid-derived compounds in S. glabra under various light conditions, allowing optimum breeding conditions to be developed for this plant.

Root morphological and physiological characteristics in maize seedlings adapted to low iron stress.

Iron (Fe) deficiency is a common challenge in crop production. Screening and research of Fe-efficient cultivars could alleviate plant stress and increase crop yields in Fe-deficient soils. In the present study, we conducted two hydroponic culture experiments with a control (100 µmol/L Fe3+-EDTA) and low Fe treatment (10 µmol/L Fe3+-EDTA) to study the morphological and physiological mechanisms of response to low Fe stress in maize hybrids seedlings. In the first experiment, we investigated 32 major maize hybrids in Southwest China. We found that six of them, including Zhenghong 2 (ZH 2), were Fe-efficient. Fifteen other cultivars, such as Chuandan 418 (CD 418), were Fe-inefficient. In the second experiment, we investigated the Fe-efficient ZH 2 and Fe-inefficient CD 418 cultivars and found that low Fe stress resulted in significant decreases in root volume, root length, number of root tips, root surface area, and root dry weight, and increased root to shoot ratio, average root diameter, and Fe-dissolution ability per mass of roots in both maize cultivars. However, the increase in Fe-dissolution ability per mass of roots in ZH 2 was higher than that in CD 418, whereas for the other measurements, the low Fe stress-induced changes in ZH 2 were less pronounced than in CD 418. Therefore, under low Fe stress, the above-mentioned growth factors in ZH 2 were higher by 54.84%, 121.46%, 107.67%, 83.96%, 140.00%, and 18.16%, respectively, than those in CD 418. In addition, leaf area, chlorophyll content, net photosynthetic rate, soluble protein content, and Catalase (CAT) and Peroxidase (POD) activities in ZH 2 were higher by 274.95%, 113.95%, 223.60%, 56.04%, 17.01% and 21.13% than those in CD 418. Therefore, compared with the Fe-inefficient cultivar (CD 418), the Fe-efficient cultivar (ZH 2) had a more developed root system and greater Fe absorption capacity per mass of roots under low iron stress, promoted the efficient absorption of Fe, maintained a higher photosynthetic area and photosynthetic rate, thereby facilitating the accumulation of photosynthetic products. Moreover, higher soluble protein content and activities of CAT and POD permitted high osmotic regulation and scavenging ability, which is an important physiological mechanism for ZH 2 adaptation to low Fe stress.

Morpho-physiological and proteomic responses to water stress in two contrasting tobacco varieties.

To gain insight into the molecular mechanisms underpinning tobacco (Nicotiana tabacum) tolerance to drought stress, we integrated anatomical, physiological, and proteomic analyses of drought-tolerant (Yuyan6, [Y6]) and -sensitive (Yunyan87 [Y87]) varieties. In comparison to Y87, Y6 exhibited higher water retention capability, improved photosynthetic performance, delayed leaf-senescence, stable leaf ultrastructure, a stronger antioxidant defense, and lesser ROS accumulation when subjected to water stress. Using an iTRAQ-based proteomics approach, 405 and 1,560 differentially accumulated proteins (DAPs) were identified from Y6 and Y87 plants, respectively, of which 114 were found to be present in both cultivars. A subsequent functional characterization analysis revealed that these DAPs were significantly enriched in eight biological processes, six molecular functions, and six cellular components and displayed differential expression patterns in Y6 and Y87 plants, suggesting that the response to water stress between both varieties differed at the proteomic level. Furthermore, we constructed protein coexpression networks and identified hub proteins regulating tobacco defenses to water stress. Additionally, qPCR analysis indicated that the majority of genes encoding selected proteins showed consistency between mRNA levels and their corresponding protein expression levels. Our results provide new insights into the genetic regulatory mechanisms associated with drought response in tobacco plants.

Tomato seeds pretreated with Antifreeze protein type I (AFP I) promotes the germination under cold stress by regulating the genes involved in germination process.

This study was conducted to investigate the involvement of antifreeze proteins (AFPs; type I and III) in the germination mechanism of tomato seeds under low temperature stress. Germination of the seeds grown at a room temperature (25°C) was observed on 5 days after sowing (DAS), while all seeds exposed to a low temperature started to germinate at 16 days after sowing (DAS). However, in comparison with control seeds (0 µg/l), seeds treated with AFP I (100, 300, or 500 µg/l) germinated earlier and at a higher percentage until 20 DAS, and seeds treated with 100 µg/l AFP I showed the highest percentage of germination. Surprisingly, AFP III did not significantly increase germination, and the rate was lower among 500 µg/l AFP III-treated seeds compared with control seeds (0 µg/l). The transcription levels of the plasma membrane-associated H+-ATPase gene and antioxidant-related superoxide dismutase (SOD) and catalase 1 (CAT1) genes were analyzed, and the transcription levels of the genes in the seeds grown at 25°C were relatively low. For low temperature-treated seeds, H+-ATPase in control seeds (0 µg/l) was higher compared with that in AFP I-treated seeds and was lower compared with that in AFP III-treated seeds. The expression levels of the antioxidant-related genes (SOD and CAT1) were lower in AFP I-treated seeds than in control seeds (0 µg/l); however, they were higher in AFP III-treated seeds than in control seeds (0 µg/l). Overall, compared with AFP III, AFP I may potentially function as a cold-protective agent by modulating the genes associated with seed germination.

Selenium downregulates auxin and ethylene biosynthesis in rice seedlings to modify primary metabolism and root architecture.

MAIN CONCLUSION: Selenium modulates the formation of primary and lateral roots through alterations in auxin and ethylene, leading to new patterns of root architecture in rice seedlings. Selenium (Se) at low concentrations can control root growth through interaction with hormone biosynthesis. Auxin and ethylene have been shown to control the root architecture, with most of the information obtained from the eudicots such Arabidopsis and Nicotiana tabacum. Here, we presented the effects of Se on auxin and ethylene pathways and examined their impact on primary metabolism and root system architecture in rice (Oryza sativa L.) seedlings. Se treatment increased elongation of primary root, but decreased the number and length of lateral roots. Se led to decreased expression of genes associated with the biosynthesis of auxin and ethylene, concomitantly with reduced production of these hormones by the roots. Moreover, Se decreased the abundance of transcripts encoding auxin transport proteins. Indole-3-acetic acid (IAA) treatment overrode the repressive effect of Se on lateral root growth. The ethylene synthesis inhibitor L-α-(2-aminoethoxyvinyl)-glycine (AVG) increased elongation of primary root, whereas the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) resulted in the opposite effect. Soluble sugars accumulate in roots of rice seedlings under Se treatment. Thus, Se modulates the formation of primary and lateral roots through alterations in auxin and ethylene, leading to new patterns of root architecture in rice seedlings.

Deciphering the nitric oxide, cyanide and iron-mediated actions of sodium nitroprusside in cotyledons of salt stressed sunflower seedlings.

Nitric oxide (NO) is an endogenous signaling molecule in plants. Sodium nitroprusside (SNP), an established NO donor used in plant science research, simultaneously releases NO, cyanide (CN-) and iron (Fe) in solution. Since cyanide and iron mask NO effect of SNP, its use in NO research is debatable. Deciphering the action of SNP through NO, CN- or Fe has been undertaken in the present work. Cotyledons from salt stressed sunflower seedlings grown in the presence of NO donors were subjected to spectrofluorometric analysis of NO, CN- and Fe contents, and proteome and biochemical analyses. Diethylenetriamine NONOate (DETA) proved to be a better NO source since SNP enhanced ROS accumulation in the tissue. Abundance of 127 proteins is modulated by salt stress. SNP and exhausted SNP (exSNP) alter the abundance of 117 and 129 proteins, respectively. These proteins belong to primary metabolism, stress-response, transport, translation, proteolysis, chaperone, regulatory, and storage. Salt-responsive proteins, such as, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK) and isocitrate dehydrogenase are negatively modulated. DETA and SNP lower the activities of GAPDH and S-adenosylmethionine synthase (SAMS). Abundance of heat shock 70 kDa protein and actin are sensitive to both NaCl and NO. SNP affects plant growth by modulating proteome though iron, cyanide and NO. Its use only as an NO donor is thus debatable. exSNP control also releases substantial amount of cyanide and iron, thus questioning its use as control in NO research.

The passion fruit liana (Passiflora edulis Sims, Passifloraceae) is tolerant to ozone.

Passiflora edulis Sims is a liana species of high economic interest and is an interesting model plant for understanding ozone action on disturbed vegetation. In this work we hypothesized that P. edulis has adaptive responses to oxidative stress that enable it to tolerate ozone damage based on its capacity to grow under a diversity of environmental conditions and to dominate disturbed areas. We exposed seedlings to three levels of ozone in a Free-Air Controlled Exposure (FACE) system (22, 41 and 58 ppb h AOT40 and 13.52, 17.24 and 20.62 mmol m-2 POD0, over 97 days) for identifying its tolerance mechanisms. Anatomical (leaf blade structure and fluorescence emission of chloroplast metabolites), physiological (leaf gas exchange, growth rate and biomass production) and biochemical (pigments, total sugars, starch, enzymatic and non-enzymatic antioxidant metabolites, reactive oxygen species and lipid peroxidation derivatives) responses were assessed. Ozone caused decreased total number of leaves, hyperplasia and hypertrophy of the mesophyll cells, and accelerated leaf senescence. However, O3 did not affect carbohydrates content, net photosynthetic rate, or total biomass production, indicating that the carboxylation efficiency and associated physiological processes were not affected. In addition, P. edulis showed higher leaf contents of ascorbic acid, glutathione (as well high ratio between their reduced and total forms), carotenoids, and flavonoids located in the chloroplast outer envelope membrane. Our results indicate that P. edulis is an O3-tolerant species due to morphological acclimation responses and an effective antioxidant defense system represented by non-enzymatic antioxidants, which maintained the cellular redox balance under ozone.

Natural selection and neutral evolutionary processes contribute to genetic divergence in leaf traits across a precipitation gradient in the tropical oak Quercus oleoides.

The impacts of drought are expanding worldwide as a consequence of climate change. However, there is still little knowledge of how species respond to long-term selection in seasonally dry ecosystems. In this study, we used QST -FST comparisons to investigate (i) the role of natural selection on population genetic differentiation for a set of functional traits related to drought resistance in the seasonally dry tropical oak Quercus oleoides and (ii) the influence of water availability at the site of population origin and in experimental treatments on patterns of trait divergence. We conducted a thorough phenotypic characterization of 1912 seedlings from ten populations growing in field and greenhouse common gardens under replicated watering treatments. We also genotyped 218 individuals from the same set of populations using eleven nuclear microsatellites. QST distributions for leaf lamina area, specific leaf area, leaf thickness and stomatal pore index were higher than FST distribution. Results were consistent across growth environments. Genetic differentiation among populations for these functional traits was associated with the index of moisture at the origin of the populations. Together, our results suggest that drought is an important selective agent for Q. oleoides and that differences in length and severity of the dry season have driven the evolution of genetic differences in functional traits.

Cerium Improves Growth of Maize Seedlings via Alleviating Morphological Structure and Oxidative Damages of Leaf under Different Stresses.

It had been indicated that cerium (Ce) could promote maize growth involving photosynthetic improvement under potassium (K) deficiency, salt stress, and combined stress of K+ deficiency and salt stress. However, whether the improved growth is related to leaf morphological structure, oxidative stress in maize leaves is not well understood. The present study showed that K+ deficiency, salt stress, and their combined stress inhibited growth of maize seedlings, affecting the formation of appendages of leaf epidermal cells, and stomatal opening, which may be due to increases in H2O2 and malondialdehyde levels, and reductions in Ca2+ content, ratios of glutathione/oxidized glutathione, ascorbic acid/dehydroascorbic acid, and the activities of superoxide dismutase, catalase, ascorbic acid peroxidase, guaiacol peroxidase, and glutathione reductase in leaves under different stresses. The adverse effects caused by combined stress were higher than those of single stress. Furthermore, our findings demonstrated that adding Ce3+ could significantly promote seedling growth, and alleviate morphological and structural damage of leaf, decrease oxidative stress and increase antioxidative capacity in maize leaves caused by different stresses.

Growth responses and accumulation of soluble sugars in Inga marginata Wild. (Fabaceae) subjected to flooding under contrasting light conditions / Respostas de crescimento e acúmulo de açúcares solúveis em Inga marginata Willd. (Fabaceae) submetida ao alagamento e sob condições contrastantes de luminosidade

Abstract Flood events in riparian forests of southern Brazil, can be characterized as unpredictable and of low magnitude with an average duration of less than 15 days. Inga marginata is an evergreen tree which grows in Southeast South America on a wide range of environments, including riparian forests. In this paper, the interactive effects of the light environment and soil flooding on morphological parameters of I. marginata were examined. Seedlings were acclimated in two contrasting light conditions: sun or shade for 30 days. Sun and shade plants were subjected to soil flooding for two periods; five or 15 days. After 5 days, the interaction between flooding and light did not affect growth, chlorophyll content and dry mass or the root-shoot ratio. After 15 days, flooded plants from the sunny treatment had a lower shoot dry mass compared to control sun plants and flooded plants from the shaded treatment. Moreover, the higher dry mass observed for shade plants compared to sun plants, following flooding, can also be directly associated with a higher content of soluble sugars. Shade plants of I. marginata showed a greater acclimation to soil waterlogging. This acclimation appears to be associated with a larger accumulation of soluble sugars compared to non-flooded plants. The responses observed on the shade plants appear to be decisive to indicate the use of I. marginata in degraded areas.

Resumo As inundações em florestas ripárias do Sul do Brasil, podem ser caracterizadas como imprevisíveis e de baixa magnitude com uma duração média de menos de 15 dias. Inga marginata é uma árvore que cresce no sudeste da América do Sul em uma grande variedade de ambientes, incluindo matas ciliares. Neste trabalho, os efeitos combinados da luminosidade e do e alagamento do solo nos parâmetros morfológicos de I. marginata foram examinados. As plântulas foram aclimatadas em duas condições contrastantes de luminosidade: sol e sombra por 30 dias. Plantas de sol e sombra foram submetidas ao alagamento do solo por dois períodos; cinco ou 15 dias. Após 5 dias, a interação entre a inundação e luminosidade não afetou o crescimento, teor de clorofila e massa seca e a razão raiz-parte aérea. Após 15 dias, plantas de sol sob inundação apresentaram menor massa seca na parte aérea em relação as plantas controle de sol e as plantas alagadas de sombra. Além disso, a maior massa seca observada nas plantas de sombra em comparação com plantas de sol, sob inundação, pode ser diretamente associado com um maior teor de açúcares solúveis. Plantas de sombra de I. marginata mostraram uma maior aclimatação ao encharcamento do solo. Esta aclimatação parece estar associada com um maior acúmulo de açúcares solúveis em comparação com as plantas não-inundadas. As respostas observadas nas plantas de sombra parecem ser determinantes para a indicação do uso de I. marginata em áreas degradadas.

Tetraploidy enhances the ability to exclude chloride from leaves in carrizo citrange seedlings.

Tetraploid citrus seedlings are more tolerant to salt stress than diploid genotypes. To provide insight into the causes of differences in salt tolerance due to ploidy and thus to better understand Cl- exclusion mechanisms in citrus, diploid and tetraploid seedlings of Carrizo citrange (CC) were grown at 0 (control) and 40mM NaCl (salt-treated) medium for 20 days. Chloride uptake and root-to-shoot translocation rates were on average 1.4-fold higher in diploid than in tetraploid salt-treated plants, which resulted in a greater (1.6-fold) Cl- build up in the leaves of the former. Root hydraulic conductance and leaf transpiration rate were 58% and 17% lower, respectively, in tetraploid than in diploid control plants. Differences remained after salt treatment which reduced these parameters by 30-40% in both genotypes. Morphology of the root system was significantly influenced by ploidy. Tetraploid roots were less branched and with lower number of root tips than those of diploid plants. The cross-section diameter and area were lower in the diploid, and consequently specific root length was higher (1.7-fold) than in tetraploid plants. The exodermis in sections close to the root apex was broader and with higher deposition of suberin in cell walls in the tetraploid than in the diploid genotype. Net CO2 assimilation rate in tetraploid salt-treated seedlings was 1.5-fold higher than in diploid salt-treated plants, likely due to the loss of photosynthetic capacity of diploid plants induced by Cl- toxicity. Leaf damage was much higher, in terms of burnt area and defoliation, in diploid than in tetraploid salt-treated plants (8- and 6-fold, respectively). Salt treatment significantly reduced (37%) the dry weight of the diploid plants, but did not affect the tetraploids. In conclusion, tetraploid CC plants appear more tolerant to salinization and this effect seems mainly due to differences in morphological and histological traits of roots affecting hydraulic conductance and transpiration rate. These results may suggest that tetraploid CC used as rootstock could improve salt tolerance in citrus trees.

Association between biometric characteristics of tomato seeds and seedling growth and development

Background The size and weight of tomato seeds depend on genetics and can be modified by environment and management. In some species, a strong relation has been described between physical aspects of the seeds and the quality of the corresponding seedlings, but this cannot be considered a general rule. The objective of this research was to identify any association between the biometric characteristics of tomato seeds and the growth and development of their seedlings. Results A total of 18 lots of hybrid tomato seeds were used (from indeterminate plants with round fruits), belonging to six varieties from two reproduction seasons. Each lot was evaluated for seed size and weight, and seed quality, in terms of the germination test (5 and 14 d after sowing). The number of normal roots emerged with a length above 2 mm was also evaluated at d 3, 4 and 5 after sowing. The length of the seedlings and their total and partial dry weight were measured 5 d after sowing. The results indicate that there was no association between seed size and weight and subsequent seedling emergence, and only weak correlations were found between the dry weight of the radicle and cotyledon and seed size. Conclusion There is little association between the physical characteristics of the seeds and the subsequent seedlings, making it impossible to propose the use of seed weight or size as a compliment to quality evaluation tests.

Estudo do potencial fitotóxico de extratos de Bauhinia ungulata L. sobre a divisão celular e atividade enzimática em plântulas de alface / Study of phytotoxic potential of extracts of Bauhinia ungulata L. on cell division and enzyme activity in lettuce seedlings

RESUMO : Este trabalho visou avaliar a ação fitotóxica de extrato e frações obtidas das folhas de Bauhinia ungulata L sobre a divisão celular e atividade das enzimas α-amilase, catalase, peroxidase e polifenoloxidase em plântulas de alface (Lactuca sativa, var Grand rapis). Sementes de alface foram expostas às concentrações de 250, 500 e 1000 µg.mL-1 do extrato de Bauhinia ungulata e frações do mesmo, em condições ambientais controladas. Após três dias de protusão radicular, as raízes das plântulas foram cortadas e submetidas a preparo específico para visualização e contagem do número de células em cada fase mitótica. No sétimo dia, as amostras foram submetidas à avaliação da atividade enzimática utilizando técnica específica para cada enzima e leitura espectrofotométrica. Na análise da divisão celular observou-se que houve redução no número de células em mitose nas raízes das plântulas testadas, e a fração acetato de etila apresentou maior efeito inibitório sobre a divisão celular. O efeito sobre as enzimas que demonstram alterações no sistema de defesa antioxidante de L. sativa mostram que o extrato bruto e frações alteraram a produção das enzimas α- amilase, peroxidase, catalase e polifenol oxidase em, ao menos, uma das concentrações testadas, sendo um indicativo de estresse que interfere na divisão celular. Os resultados obtidos apontam para a presença de compostos com atividade inibidora ou estimulatória nas folhas de B. ungulata revelando potencial fitotóxico sobre as plântulas de alface

ABSTRACT Study of phytotoxic potential of extracts of Bauhinia ungulata L. on cell division and enzyme activity in lettuce seedlings. Theaim of this studywas to evaluate thephytotoxic actionofextractand fractionsobtainedfrom the leaves ofBauhiniaungulata L. inbioassaylaboratory, assessing their interferenceon cell division andactivity of the enzymesalpha-amylase, catalase, peroxidase andpolyphenoloxidaseofLactucasativa(lettu ce). The lettuce seedswere exposedto concentrationsof 250,500and 1000µg.mL-1of the extracts and fractions of Bauhinia ungulataunder controlled environmental conditions. After three days ofrootradicleprotrusion, the plants` rootswere cutand subjected tospecific preparationfor visualization andcounting of the number ofcells in eachmitotic stage. On the seventh daythe sampleswere subjected toenzymatic activityusingspecific techniquefor eachenzyme anda spectrophotometer measure. In the analysisof the mitotic indexit was observedthatthere was reduction in the number of cells in mitosis in the roots of the plants tested and thattheethyl acetate fractionsignificantly affected thecell division. The effecton the enzymesthat showed changesinthe antioxidant defensesystemof Lactucasativaindicate thatthe crude extractand the fractionsaltered theproductionofα-amylase, peroxides, catalaseand polyphenol enzymesin at least oneof the testedconcentrations, representingan indicationstress,which interferesin the celldivision.Theobtained results point out the presence ofcompoundswithinhibitoryorstimulatory activityon plant leaves of B. ungulata,revealingphytotoxic potentialtolettuce seedlings

Salinity stress constrains photosynthesis in Fraxinus ornus more when growing in partial shading than in full sunlight: consequences for the antioxidant defence system.

BACKGROUND AND AIMS: A major challenge in plant ecophysiology is understanding the effects of multiple sub-optimal environmental conditions on plant performance. In most Mediterranean areas soil salinity builds up during the summer because of low availability of soil water coupled with hot temperatures. Although sunlight and soil salinity may strongly interact in determining a plant's performance, this has received relatively little attention. METHODS: Two-year-old seedlings of Fraxinus ornus were grown outdoors in pots during a Mediterranean summer in either 45 % (shaded plants) or 100 % (sun plants) sunlight irradiance and were supplied with either deionized water or deionized water plus 75 mm NaCl. Morpho-anatomical traits, water and ionic relations, gas exchange and photosystem II performance, concentrations of individual carotenoids, activity of antioxidant enzymes, concentrations of ascorbic acid and individual polyphenols were measured in leaves. Leaf oxidative stress and damage were estimated by in vivo analysis of stable free radicals and ultrastructural analyses. KEY RESULTS: Leaf concentrations of potentially toxic ions did not markedly differ in shaded or sun plants in response to salinity. Leaves of sun plants displayed superior water use efficiency compared with leaves of shaded plants, irrespective of salinity treatment, and had both better stomatal control and higher CO2 carboxylation efficiency than leaves of shaded plants. In the salt-treated groups, the adverse effects of excess midday irradiance were greater in shade than in sun plants. The activity of enzymes responsible for detoxifying hydrogen peroxide decreased in shaded plants and increased in sun plants as a result of salinity stress. In contrast, the activity of guaiacol peroxidase and the concentration of phenylpropanoids increased steeply in response to salinity in shaded plants but were unaffected in sun plants. CONCLUSIONS: It is concluded that salinity may constrain the performance of plants growing under partial shading more severely than that of plants growing under full sun during summer. The results suggest co-ordination within the antioxidant defence network aimed at detoxifying salt-induced generation of reactive oxygen species.

Effect of crude oil contamination on the chlorophyll content and morpho-anatomy of Cyperus brevifolius (Rottb.) Hassk.

Chlorophyll plays a pivotal role in the plant physiology and its productivity. Cultivation of plants in crude oil contaminated soil has a great impact on the synthesis of chlorophyll pigment. Morpho-anatomy of the experimental plant also shows structural deformation in higher concentrations. Keeping this in mind, a laboratory investigation has been carried out to study the effect of crude oil on chlorophyll content and morpho-anatomy of Cyperus brevifolius plant. Fifteen-day-old seedling of the plant was planted in different concentrations of the crude oil mixed soil (i.e., 10,000, 20,000, 30,000, 40,000, and 50,000 ppm). A control setup was also maintained without adding crude oil. Results were recorded after 6 months of plantation. Investigation revealed that there is a great impact of crude oil contamination on chlorophyll content of the leaves of the experimental plant. It also showed that chlorophyll a, chlorophyll b, and total chlorophyll content of leaves grown in different concentrations of crude oil were found to be lower than those of the control plant. Further, results also demonstrated that chlorophyll content was lowest in the treatment that received maximum dose of crude oil. It also showed that chlorophyll content was decreased with increased concentration of crude oil. Results also demonstrated that there was a reduction in plant shoot and root biomass with the increase of crude oil concentration. Results also revealed that the shoot biomass is higher than root biomass. Morphology and anatomy of the experimental plant also show structural deformation in higher concentrations. Accumulation of crude oil on the cuticle of the transverse section of the leaves and shoot forms a thick dark layer. Estimation of the level of pollution in an environment due to oil spill is possible by the in-depth study of the harmful effects of oil on the morphology and anatomy and chlorophyll content of the plants grown in that particular environment.

Impact of TiO2 nanoparticles on Vicia narbonensis L.: potential toxicity effects.

This work was aimed to provide further information about toxicology of TiO2 nanoparticles (NPs) on Vicia narbonensis L., considering different endpoints. After exposure to TiO2 nanoparticle suspension (mixture of rutile and anatase, size <100 nm) at four different concentrations (0.2, 1.0, 2.0 and 4.0 ‰), the seeds of V. narbonensis were let to germinate in controlled environmental conditions. After 72 h, the extent of the success of the whole process (seed germination plus root elongation) was recorded as the vigour index, an indicator of possible phytotoxicity. After the characterisation of the hydric state of different materials, oxidative stress and enzymatic and nonenzymatic antioxidant responses were considered as indicators of possible cytotoxicity and to assess if damage induced by TiO2 NPs was oxidative stress-dependent. Cytohistochemical detection of in situ DNA fragmentation as genotoxicity endpoint was monitored by TUNEL reaction. The treatments with TiO2 NPs in our system induced phytotoxic effects, ROS production and DNA fragmentation. The nonenzymatic and enzymatic antioxidant responses were gradually and differentially activated and were able to maintain the oxidative damage to levels not significantly different from the control. On the other hand, the results of DNA fragmentation suggested that the mechanisms of DNA repair were not effective enough to eliminate early genotoxicity effects.

Acropetal leaflet initiation of Eschscholzia californica is achieved by constant spacing of leaflets and differential growth of leaf.

In compound leaves, leaflet primordia are initiated directionally along the lateral sides. Our understanding of the molecular basis of leaflet initiation has improved, but the regulatory mechanisms underlying spatio-temporal patterns remain unclear. In this study, we investigated the mechanisms of acropetal (from the base to the tip) progression of leaflet initiation in Eschscholzia californica. We established an ultraviolet-laser ablation system to manipulate compound-leaf development. Local ablation at the leaflet incipient site generated leaves with asymmetric morphology. In the majority of cases, leaflets that were initiated on the ablated sides shifted apically. Finite time-course observation revealed that the timing of leaflet initiation was delayed, but the distance from the leaf tip did not decrease. These results were suggestive of the local spacing mechanism in leaflet initiation, whereby the distance from the leaf tip and adjacent pre-existing leaflet determines the position of leaflet initiation. To understand how such a local patterning mechanism generates a global pattern of successive leaflet initiation, we assessed the growth rate gradient along the apical-basal axis. Our time-course analysis revealed differential growth rates along the apical-basal axis of the leaf, which can explain the acropetal progression of leaflet initiation. We propose that a leaflet is initiated at a site where the distances from pre-existing leaflets and the leaf tip are sufficient. Furthermore, the differential growth rate may be a developmental factor underlying the directionality of leaflet initiation.