Physiological and metabolic alterations in basil (Ocimum basilicum L) varieties under distinct soil water levels / Alteraciones fisiológicas y metabólicas en variedades de albahaca (Ocimum basilicum L) bajo distintos niveles de agua del suelo

Basil (Ocimum basilicumL.) is a medicinal species used in several areas, such as food, medicines and cosmetics, and the understanding of its physiological behavior under environmental conditions is of paramount importance for the improvement of cultivation methods. The objective of this study was to evaluate the influence of different water availability under physiological, biochemical and metabolic characteristics, in three distinct genotypes: 'Alfavaca basilicão', 'Gennaro de menta' and 'Grecco à palla', during two different phenological stages (vegetative and reproductive). It was found that the water deficit promotes physiological changes to tolerate water stress, and the studied genotypes have different routes to achieve this physiological tolerance, which culminates in a distinct accumulation of metabolites in plants, and can be considered interesting if the final product is the production of essential oils.

La albahaca (Ocimum basilicum L.) es una planta medicinal utilizada en varias áreas: alimenticia, medicinal e industria cosmética; es de suma importancia el entendimiento de su comportamiento fisiológico bajo diferentes condiciones ambientales con el fin de mejorar los procesos del cultivo. El objetivo de este estudio fue evaluar la influencia de diferentes disponibilidades hídricas en las características fisiológicas, bioquímicas y metabólicas en tres genotipos de albahaca: "Alfavaca basilicão", "Gennaro de menta" y "Grecco à palla" durante dos etapas fenológicas (vegetativa y reproductiva). Fue encontrado que el déficit hídrico promueve cambios fisiológicos con el fin de tolerar el estrés hídrico. Los genotipos estudiados presentaron diferentes rutas para alcanzar esta tolerancia fisiológica, la cual culmina con distintas acumulaciones de metabolitos en las plantas, y puede ser considerado interesante si el producto final es la producción de aceites esenciales.

Development of novel robotic platforms for mechanical stress induction, and their effects on plant morphology, elements, and metabolism.

This research evaluates the effect on herbal crops of mechanical stress induced by two specially developed robotic platforms. The changes in plant morphology, metabolite profiles, and element content are evaluated in a series of three empirical experiments, conducted in greenhouse and CNC growing bed conditions, for the case of basil plant growth. Results show significant changes in morphological features, including shortening of overall stem length by up to 40% and inter-node distances by up to 80%, for plants treated with a robotic mechanical stress-induction protocol, compared to control groups. Treated plants showed a significant increase in element absorption, by 20-250% compared to controls, and changes in the metabolite profiles suggested an improvement in plants' nutritional profiles. These results suggest that repetitive, robotic, mechanical stimuli could be potentially beneficial for plants' nutritional and taste properties, and could be performed with no human intervention (and therefore labor cost). The changes in morphological aspects of the plant could potentially replace practices involving chemical treatment of the plants, leading to more sustainable crop production.

Sweet Basil Has Distinct Synthases for Eugenol Biosynthesis in Glandular Trichomes and Roots with Different Regulatory Mechanisms.

Production of a volatile phenylpropene; eugenol in sweet basil is mostly associated with peltate glandular trichomes (PGTs) found aerially. Currently only one eugenol synthase (EGS), ObEGS1 which belongs to PIP family is identified from sweet basil PGTs. Reports of the presence of eugenol in roots led us to analyse other EGSs in roots. We screened for all the PIP family reductase transcripts from the RNA-Seq data. In vivo functional characterization of all the genes in E. coli showed their ability to produce eugenol and were termed as ObEGS2-8. Among all, ObEGS1 displayed highest expression in PGTs and ObEGS4 in roots. Further, eugenol was produced only in the roots of soil-grown plants, but not in roots of aseptically-grown plants. Interestingly, eugenol production could be induced in roots of aseptically-grown plants under elicitation suggesting that eugenol production might occur as a result of environmental cues in roots. The presence of ObEGS4 transcript and protein in aseptically-grown plants indicated towards post-translational modifications (PTMs) of ObEGS4. Bioinformatics analysis showed possibility of phosphorylation in ObEGS4 which was further confirmed by in vitro experiment. Our study reveals the presence of multiple eugenol synthases in sweet basil and provides new insights into their diversity and tissue specific regulation.

Modified multiwall carbon nanotubes display either phytotoxic or growth promoting and stress protecting activity in Ocimum basilicum L. in a concentration-dependent manner.

Carbon-based materials including multiwall carbon nanotubes (MWCNTs) have been recently implicated in a number of reports dealing with their potential use in agriculture, leading to contradictory findings. In this study, MWCNTs were successfully functionalized with carboxylic acid groups (MWCNTs-COOH) in order to increase water dispersion. Hydroponically cultured sweet basil (Ocimum basilicum L.) seedlings were subjected to four concentrations (0, 25, 50 and 100 mg L-1) of MWCNTs-COOH under three salt stress levels (0, 50 and 100 mM NaCl). An array of agronomic, physiological, analytical and biochemical parameters were evaluated in an attempt to examine the potential use of MWCNTs in plants under optimal and abiotic stress conditions. Application of MWCNTs-COOH at optimum concentration (50 mg L-1) could ameliorate the negative effects of salinity stress by increasing chlorophyll and carotenoids content and inducing non-enzymatic (i.e. phenolic content) and enzymatic antioxidant components (i.e. ascorbate peroxidase (APX), catalase (CAT) and guaiacol peroxidase (GP) activity). Furthermore, MWCNTs-COOH treatments under optimal conditions induced plant growth, while a significant increase (P ≤ 0.01) was recorded in essential oil content and compound profile. On the other hand, biochemical and epifluorescence microscopy evidence suggested that high dosage (100 mg L-1) of MWCNTs-COOH leads to toxicity effects in plant tissue. Overall, the positive response of plants to low concentrations of MWCNTs-COOH under control and abiotic stress conditions renders them as potential novel plant growth promoting and stress protecting agents, opening up new perspectives for their use in agriculture.

"Help is in the air": volatiles from salt-stressed plants increase the reproductive success of receivers under salinity.

MAIN CONCLUSION: Salinity alters VOC profile in emitter sweet basil plants. Airborne signals by emitter plants promote earlier flowering of receivers and increase their reproductive success under salinity. Airborne signals can prime neighboring plants against pathogen and/or herbivore attacks, whilst little is known about the possibility that volatile organic compounds (VOCs) emitted by stressed plants alert neighboring plants against abiotic stressors. Salt stress (50 mM NaCl) was imposed on Ocimum basilicum L. plants (emitters, namely NaCl), and a putative alerting-priming interaction was tested on neighboring basil plants (receivers, namely NaCl-S). Compared with the receivers, the NaCl plants exhibited reduced biomass, lower photosynthesis, and changes in the VOC profile, which are common early responses of plants to salinity. In contrast, NaCl-S plants had physiological parameters similar to those of nonsalted plants (C), but exhibited a different VOC fingerprint, which overlapped, for most compounds, with that of emitters. NaCl-S plants exposed later to NaCl treatment (namely NaCl-S + NaCl) exhibited changes in the VOC profile, earlier plant senescence, earlier flowering, and higher seed yield than C + NaCl plants. This experiment offers the evidence that (1) NaCl-triggered VOCs promote metabolic changes in NaCl-S plants, which, finally, increase reproductive success and (2) the differences in VOC profiles observed between emitters and receivers subjected to salinity raise the question whether the receivers are able to "propagate" the warning signal triggered by VOCs in neighboring companions.

Interaction between hydrogen peroxide and sodium nitroprusside following chemical priming of Ocimum basilicum L. against salt stress.

Sodium nitroprusside (SNP) and hydrogen peroxide (H2 O2 ), as priming agents, have the well-recorded property to increase plant tolerance against a range of different abiotic stresses such as salinity. In this regard, the present study was conducted to evaluate the effect of different levels of SNP (100 and 200 µM) and H2 O2 (2.5 and 5 mM) as well as their combinations under salt stress (0 and 50 mM NaCl) on key physiological and biochemical attributes of the economically important aromatic plant basil (Ocimum basilicum L.) grown under hydroponic culture. Results revealed that morphological parameters such as plant height, root length, leaf fresh and dry weights (FW and DW) were significantly decreased by salinity stress, while SNP and H2 O2 treatments, alone or combined, increased FW and DW thus enhancing plant tolerance to salt stress. Furthermore, 200 µM SNP + 2.5 mM H2 O2 appeared to be the most effective treatment by causing significant increase in chlorophyll a and b, anthocyanin content and guaiacol peroxidase and ascorbate peroxidase enzymes activities under saline condition. In addition, analytical measurements showed that essential oil profile (concentration of main components) under salt stress was mostly affected by SNP and H2 O2 treatments. The highest increase was observed for methyl chavicol (43.09-69.91%), linalool (4.8-17.9%), cadinol (1.5-3.2%) and epi-α-cadinol (0.18-10.75%) compounds. In conclusion, current findings demonstrated a positive crosstalk between SNP and H2 O2 toward improved basil plant tolerance to salt stress, linked with regulation of essential oil composition.

Spectroscopy can predict key leaf traits associated with source-sink balance and carbon-nitrogen status.

Approaches that enable high-throughput, non-destructive measurement of plant traits are essential for programs seeking to improve crop yields through physiological breeding. However, many key traits still require measurement using slow, labor-intensive, and destructive approaches. We investigated the potential to retrieve key traits associated with leaf source-sink balance and carbon-nitrogen status from leaf optical properties. Structural and biochemical traits and leaf reflectance (500-2400 nm) of eight crop species were measured and used to develop predictive 'spectra-trait' models using partial least squares regression. Independent validation data demonstrated that the models achieved very high predictive power for C, N, C:N ratio, leaf mass per area, water content, and protein content (R2>0.85), good predictive capability for starch, sucrose, glucose, and free amino acids (R2=0.58-0.80), and some predictive capability for nitrate (R2=0.51) and fructose (R2=0.44). Our spectra-trait models were developed to cover the trait space associated with food or biofuel crop plants and can therefore be applied in a broad range of phenotyping studies.

Molecular cloning, functional characterization and expression of a drought inducible phenylalanine ammonia-lyase gene (ObPAL) from Ocimum basilicum L.

Phenylalanine ammonia-lyase (PAL) is a control point for branched phenylpropanoid and terpenoid pathways. It represents the first regulatory step to provide a metabolic flux to produce of the precursors needed for biosynthesizing main volatile phenylpropanoid compounds (methyleugenol and methylchavicol) in basil. It is crucial during the stage of the environmental and development stimulants. To obtain better knowledge of the biosynthesis of these phenylpropene compounds, characterization and cloning of Ocimum basilicum PAL (ObPAL) cDNA and its heterologous expression and enzyme activity were assessed. The almost full-length ObPAL was 2064 bp in size encoding a 687-amino-acid polypeptide with molecular weight of 74.642 kDa and theoretical pI of 8.62. Phylogenetic analysis revealed a significant evolutionary relatedness of ObPAL with the PAL sequence reported in different species of Lamiaceae. To further confirm its function, ObPAL was cloned into pET28a (+) vector and expressed in E. coli. The recombinant protein exhibited high PAL activity and could catalyze the L-Phe conversion to trans-cinnamic acid. Expression analysis of PAL gene showed that ObPAL manifested various transcription ratios exposed to drought stress. Overall, our results demonstrated the ObPAL regulation gene is possibly a mechanism dependent on cultivar and drought stress.

Effects of the exposure of TiO2 nanoparticles on basil (Ocimum basilicum) for two generations.

There is a lack of information about the transgenerational effects of titanium dioxide nanoparticles (nano-TiO2) in plants. This study aimed to evaluate the impacts of successive exposure of nano-TiO2 with different surface properties to basil (Ocimum basilicum). Seeds from plants exposed or re-exposed to pristine, hydrophobic, or hydrophilic nano-TiO2 were cultivated for 65 days in soil unamended or amended with 750 mg·kg-1 of the respective particles. Plant growth, concentration of titanium and essential elements, as well as content of carbohydrates and chlorophyll were evaluated. There were no differences on Ti concentration in roots of plants sequentially exposed to pristine or hydrophobic nano-TiO2, or in roots of plants exposed to the corresponding particle, only in the second cycle. However, sequential exposure to hydrophilic particles resulted in 65.2% less Ti in roots, compared to roots of plants exposed the same particles, only in the second cycle. The Ti concentrations in shoots were similar in all treatments. On the other hand, pristine and hydrophilic particles reduced Mg in root by 115% and 81%, respectively, while pristine and hydrophobic particles reduced Ni in shoot by 84% and 75%, respectively, compared to unexposed plants in both cycles. Sequential exposure to pristine nano-TiO2 increased stomatal conductance (214%, p ≤ 0.10), compared to plants that were never exposed. Hydrophobic and hydrophilic nano-TiO2 reduced chlorophyll b (52%) and total chlorophyll (30%) but increased total sugar (186%) and reducing sugar (145%), compared to unexposed plants in both cycles. Sequential exposure to hydrophobic or hydrophilic nano-TiO2 resulted in more adverse effects on photosynthesis but in positive effects on plant growth, compared to pristine nano-TiO2.

Water deficit stress fluctuates expression profiles of 4Cl, C3H, COMT, CVOMT and EOMT genes involved in the biosynthetic pathway of volatile phenylpropanoids alongside accumulation of methylchavicol and methyleugenol in different Iranian cultivars of basil.

Here, for the first time, the accumulation ratio of methylchavicol and methyleugenoland compounds together with the expression profiles of five critical genes (i.e., 4Cl, C3H, COMT, CVOMT and EOMT) in three Iranian cultivars of basil were assessed under water deficit stress at flowering stage. The highest value of methylchavicol was detected for Cul. 3 under severe stress (S3; 7.695µg/mg) alongside Cul. 2 under similar circumstances (S3; 4.133µg/mg), while regarding Cul. 1, no detectable amounts were acquired. Considering methyleugenol, Cul. 3 (0.396µg/mg; S0) followed by Cul. 1 (S3; 0.160µg/mg) were the capable plant samples in producing some detectable amounts of methyleugenol. Apart from some expectations, all the genes under study exhibited also different transcription ratios under deficit stress. Our results, overall, demonstrated that the regulation of the above-mentioned genes and production of methychavicol and methyleugenol seems to be a cultivar- and drought stress-dependent mechanism.

Changes in visual quality, physiological and biochemical parameters assessed during the postharvest storage at chilling or non-chilling temperatures of three sweet basil (Ocimum basilicum L.) cultivars.

Leaves of three different sweet basil (Ocimum basilicum L.) cultivars (Italico a foglia larga, Cammeo, and Italiano classico) packed in macro-perforated polyethylene bags were stored at chilling (4°C) or non-chilling temperature (12°C) for 9days. During storage, visual quality, physiological (respiration rate, ethylene production, ammonium content) and chemical (antioxidant activity, total polyphenols and polyphenol profile) parameters were measured. Detached leaves stored at chilling temperature showed visual symptoms related to chilling injury, while ethylene production and ammonium content resulted associated to cultivar sensibility to damage at low temperature. Storage at 4°C caused a depletion in polyphenols content and antioxidant capability, which was preserved at 12°C. Regarding the polyphenols profile, stressful storage conditions did not enhance the phenolic metabolism. However, leaves stored at 12°C did not loss a significant amount of metabolites respect to fresh leaves, suggesting the possibility to extend the storability after the expiration date, for a possible recovery of bioactive compounds.

Dissecting molecular and physiological response mechanisms to high solar radiation in cyanic and acyanic leaves: a case study on red and green basil.

Photosynthetic performance and the expression of genes involved in light signaling and the biosynthesis of isoprenoids and phenylpropanoids were analysed in green ('Tigullio', TIG) and red ('Red Rubin', RR) basil. The aim was to detect the physiological and molecular response mechanisms to high sunlight. The attenuation of blue-green light by epidermal anthocyanins was shown to evoke shade-avoidance responses with consequential effects on leaf morpho-anatomical traits and gas exchange performance. Red basil had a lower mesophyll conductance, partially compensated by the less effective control of stomatal movements, in comparison with TIG. Photosynthesis decreased more in TIG than in RR in high sunlight, because of larger stomatal limitations and the transient impairment of PSII photochemistry. The methylerythritol 4-phosphate pathway promoted above all the synthesis and de-epoxidation of violaxanthin-cycle pigments in TIG and of neoxanthin and lutein in RR. This enabled the green leaves to process the excess radiant energy effectively, and the red leaves to optimize light harvesting and photoprotection. The greater stomatal closure observed in TIG than in RR was due to enhanced abscisic acid (ABA) glucose ester deglucosylation and reduced ABA oxidation, rather than to superior de novo ABA synthesis. This study shows a strong competition between anthocyanin and flavonol biosynthesis, which occurs at the level of genes regulating the oxidation of the C2-C3 bond in the dihydro-flavonoid skeleton.

A cold-tolerant evergreen interspecific hybrid of Ocimum kilimandscharicum and Ocimum basilicum: analyzing trichomes and molecular variations.

Ocimum (Lamiaceae) is an important source of essential oils and aroma chemicals especially eugenol, methyl eugenol, linalool, methyl chavicol etc. An elite evergreen hybrid has been developed from Ocimum kilimandscharicum and Ocimum basilicum, which demonstrated adaptive behavior towards cold stress. A comparative molecular analysis has been done through RAPD, AFLP, and ISSR among O. basilicum and O. kilimandscharicum and their evergreen cold-tolerant hybrid. The RAPD and AFLP analyses demonstrated similar results, i.e., the hybrid of O. basilicum and O. kilimandscharicum shares the same cluster with O. kilimandscharicum, while O. basilicum behaves as an outgroup, whereas in ISSR analysis, the hybrid genotype grouped in the same cluster with O. basilicum. Ocimum genotypes were analyzed and compared for their trichome density. There were distinct differences on morphology, distribution, and structure between the two kinds of trichomes, i.e., glandular and non-glandular. Glandular trichomes contain essential oils, polyphenols, flavonoids, and acid polysaccharides. Hair-like trichomes, i.e., non-glandular trichomes, help in keeping the frost away from the living surface cells. O. basilicum showed less number of non-glandular trichomes on leaves compared to O. kilimandscharicum and the evergreen cold-tolerant hybrid. Trichomes were analyzed in O. kilimandscharicum, O. basilicum, and their hybrid. An increased proline content at the biochemical level represents a higher potential to survive in a stress condition like cold stress. In our analysis, the proline content is quite higher in tolerant variety O. kilimandscharicum, low in susceptible variety O. basilicum, and intermediate in the hybrid. Gene expression analysis was done in O. basilicum, O. kilimandscharicum and their hybrid for TTG1, GTL1, and STICHEL gene locus which regulates trichome development and its formation and transcription factors WRKY and MPS involved in the regulation of plant responses to freezing and cold. The analysis showed that O. kilimandscharicum and the hybrid were very close to each other but O. basilicum was more distinct in all respects. The overexpression of the WRKY coding gene showed high expression in the hybrid as compared to O. kilimandscharicum and O. basilicum and the transcription factor microspore-specific (MPS) promoter has also shown overexpression in the hybrid for its response against cold stress. The developed evergreen interspecific hybrid may thus provide a base to various industries which are dependent upon the bioactive constituents of Ocimum species.

Resistance Against Basil Downy Mildew in Ocimum Species.

Downy mildew, caused by the oomycete Peronospora belbahrii, is a devastating disease of sweet basil. In this study, 113 accessions of Ocimum species (83 Plant Introduction entries and 30 commercial entries) were tested for resistance against downy mildew at the seedling stage in growth chambers, and during three seasons, in the field. Most entries belonging to O. basilicum were highly susceptible whereas most entries belonging to O. americanum, O. kilimanadascharicum, O. gratissimum, O. campechianum, or O. tenuiflorum were highly resistant at both the seedling stage and the field. Twenty-seven highly resistant individual plants were each crossed with the susceptible sweet basil 'Peri', and the F1 progeny plants were examined for disease resistance. The F1 plants of two crosses were highly resistant, F1 plants of 24 crosses were moderately resistant, and F1 plants of one cross were susceptible, suggesting full, partial, or no dominance of the resistance gene(s), respectively. These data confirm the feasibility of producing downy mildew-resistant cultivars of sweet basil by crossing with wild Ocimum species.

Photoprotection by foliar anthocyanins mitigates effects of boron toxicity in sweet basil (Ocimum basilicum).

Boron (B) toxicity is an important agricultural problem in arid environments. Excess edaphic B compromises photosynthetic efficiency, limits growth and reduces crop yield. However, some purple-leafed cultivars of sweet basil (Ocimum basilicum) exhibit greater tolerance to high B concentrations than do green-leafed cultivars. We hypothesised that foliar anthocyanins protect basil leaf mesophyll from photo-oxidative stress when chloroplast function is compromised by B toxicity. Purple-leafed 'Red Rubin' and green-leafed 'Tigullio' cultivars, grown with high or negligible edaphic B, were given a photoinhibitory light treatment. Possible effects of photoabatement by anthocyanins were simulated by superimposing a purple polycarbonate filter on the green leaves. An ameliorative effect of light filtering on photosynthetic quantum yield and on photo-oxidative load was observed in B-stressed plants. In addition, when green protoplasts from both cultivars were treated with B and illuminated through a screen of anthocyanic protoplasts or a polycarbonate film which approximated cyanidin-3-O-glucoside optical properties, the degree of photoinhibition, hydrogen peroxide production, and malondialdehyde content were reduced. The data provide evidence that anthocyanins exert a photoprotective role in purple-leafed basil mesophyll cells, thereby contributing to improved tolerance to high B concentrations.

Epidermal coumaroyl anthocyanins protect sweet basil against excess light stress: multiple consequences of light attenuation.

The putative photoprotective role of foliar anthocyanins continues to attract heated debate. Strikingly different experimental set-ups coupled with a poor knowledge of anthocyanin identity have likely contributed to such disparate opinions. Here, the photosynthetic responses to 30 or 100% solar irradiance were compared in two cultivars of basil, the green-leafed Tigullio (TG) and the purple-leafed Red Rubin (RR). Coumaroyl anthocyanins in RR leaf epidermis significantly mitigated the effects of high light stress. In full sunlight, RR leaves displayed several shade-plant traits; they transferred less energy than did TG to photosystem II (PSII), and non-photochemical quenching was lower. The higher xanthophyll cycle activity in TG was insufficient to prevent inactivation of PSII in full sunlight. However, TG was the more efficient in the shade; RR was far less able to accommodate a large change in irradiance. Investment of carbon to phenylpropanoid biosynthesis was more in RR than in TG in the shade, and was either greatly enhanced in TG or varied little in RR because of high sunlight. The metabolic cost of photoprotection was lower whereas light-induced increase in biomass production was higher in RR than in TG, thus making purple basil the more light tolerant. Purple basil appears indeed to display the conservative resource-use strategy usually observed in highly stress tolerant species. We conclude that the presence of epidermal coumaroyl anthocyanins confers protective benefits under high light, but it is associated with a reduced plasticity to accommodate changing light fluxes as compared with green leaves.

Stomatal density and metabolic determinants mediate salt stress adaptation and water use efficiency in basil (Ocimum basilicum L.).

Increasing salinity tolerance and water-use efficiency in crop plants are two major challenges that agriculture must face in the next decades. Many physiological mechanisms and molecular components mediating crop response to environmental stresses have been identified. However, the functional inter-links between stress adaptation responses have not been completely understood. Using two basil cultivars (Napoletano and Genovese) with contrasting ability to respond to salt stress, here we demonstrate that reduced stomatal density, high ascorbate level and polyphenol oxidase (PPO) activity coordinately contribute to improve basil adaptation and water use efficiency (WUE) in saline environment. The constitutively reduced stomatal density was associated with a "delayed" accumulation of stress molecules (and growth inhibiting signals) such as abscisic acid (ABA) and proline, in the more tolerant Genovese. Leaf volatile profiling also revealed cultivar-specific patterns, which may suggest a role for the volatile phenylpropanoid eugenol and monoterpenes in conferring stress tolerance via antioxidant and signalling functions.

Superação de dormência em sementes de manjericão (Ocimum basilicum L. ) / Overcoming dormancy in seeds of basil (Ocimum basilicum L. )

O objetivo do trabalho foi avaliar a eficiência de tratamentos pré-germinativos na superação da dormência de sementes de manjericão, produzidas no Horto de Plantas Medicinais da Unimontes, em fevereiro de 2011. Foram realizadas as seguintes determinações para avaliação da qualidade fisiológica das sementes: teor de água, germinação, primeira contagem de germinação, emergência de plântulas e índice de velocidade de emergência. O delineamento experimental utilizado foi inteiramente casualizado com quatro repetições de 50 sementes por tratamento, sendo T1- testemunha; T2 - pré esfriamento das sementes em câmara tipo BOD sob temperatura de 10ºC por 4 dias; T3 - embebição das sementes em água destilada por 24 horas; T4 - embebição das sementes em solução contendo KNO3 a 0,2 % por 5 minutos e T5 - sementes submetidas em água destilada a temperatura de 70ºC por 5 minutos. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste Scott - Knott a 5% de probabilidade. O tratamento pré esfriamento em câmara tipo BOD a 10ºC por 4 dias reduz a dormência e promove incrementos na qualidade fisiológica das sementes do manjericão.

Aiming in order to assess the effectiveness of treatments to overcome dormancy in seeds of basil, an experiment was conducted at the Laboratory of Seed Analysis of Unimontes. Following determinations were performed to evaluate the physiological quality of seeds, water content, germination, first count germination, seedling emergence and emergence speed index. The experimental design was completely randomized design with four replications of 50 seeds per treatment, which consisted of: T1 - control, T2 - pre-cooling of the seed chamber BOD at a temperature of 10ºC for 4 days, T3 - soaking the seeds in water distilled for 24 hours, T4 - soaking the seeds in a solution containing 0,2% for 5 minutes and T5 - submitted seeds in distilled water at 70ºC for 5 minutes. Data were subjected to analysis of variance and the averages compared by Scott-Knott 5% probability. The pre-cooling treatment in BOD chamber at 10ºC for 4 days reduced dormancy and promotes increases in the physiological quality of seeds of basil.

Guard cell apoplastic photosynthate accumulation corresponds to a phloem-loading mechanism.

Apoplastic phloem loaders have an apoplastic step in the movement of the translocated sugar, prototypically sucrose, from the mesophyll to the companion cell-sieve tube element complex. In these plants, leaf apoplastic sucrose becomes concentrated in the guard cell wall to nominally 150 mM by transpiration during the photoperiod. This concentration of external sucrose is sufficient to diminish stomatal aperture size in an isolated system and to regulate expression of certain genes. In contrast to apoplastic phloem loaders and at the other extreme, strict symplastic phloem loaders lack an apoplastic step in phloem loading and mostly transport raffinose family oligosaccharides (RFOs), which are at low concentrations in the leaf apoplast. Here, the effects of the phloem-loading mechanism and associated phenomena on the immediate environment of guard cells are reported. As a first step, carbohydrate analyses of phloem exudates confirmed basil (Ocimum basilicum L. cv. Minimum) as a symplastic phloem-loading species. Then, aspects of stomatal physiology of basil were characterized to establish this plant as a symplastic phloem-loading model species for guard cell research. [(14)C]Mannitol fed via the cut petiole accumulated around guard cells, indicating a continuous leaf apoplast. The (RFO+sucrose+hexoses) concentrations in the leaf apoplast were low, <0.3 mM. Neither RFOs (<10 mM), sucrose, nor hexoses (all, P >0.2) were detectable in the guard cell wall. Thus, differences in phloem-loading mechanisms predict differences in the in planta regulatory environment of guard cells.

The effect of 4-nonylphenol on the pigmentation of Ocimum basilicum (Basil).

BACKGROUND, AIM AND SCOPE: Tests during the last few years have confirmed that 4-nonylphenol (4-NP) can have oestrogen-like effects (xeno-hormone) on animal organisms. The objective was to firstly evaluate the ecotoxicological effects of 4-NP on plants such as hydrocultures. To clarify how this substance interferes with the photosynthetic system of plants, various tests were carried out using the basil plant (Ocimum basilicum). METHODS: The effect of the pollutant 4-NP on the pigment content in the leaves of the basil plant was analysed with the use of High-Performance-Liquid-Chromatography (HPLC). RESULTS AND DISCUSSION: A general assessment of the HPLC data revealed that plants that came in contact with the 4-nonylphenol showed a change in pigmentation. More chlorophyll a and b was produced, although at the same time a higher production of degradation products and by-products of the chlorophylls was observed. These occurrences can therefore be seen as an impairment of the photosynthetic process. The contaminated plants produced less xanthophylls than the non-contaminated ones, though these differences were statistically not significant. CONCLUSION AND PERSPECTIVE: The variations on the pigment content in the leaves of the basil plant can be interpreted as a consequence of the 4-NP application. It was, however, not investigated whether the plants absorbed the pollutants directly. The effect could have been caused by adsorption of the oily substance to the roots, and this could have led to a hindrance of the uptake of nutrients and possibly water. In order to clarify this further, biochemical experiments are being conducted.