Bioprospecting beneficial endophytic bacterial communities associated with Rosmarinus officinalis for sustaining plant health and productivity.

The present study aimed to isolate and identify root endophytic bacteria with multifunctional plant growth promoting (PGP) traits from medicinal plant Rosmarinus officinalis grown in the North-Western Himalayas. A total of 42 strains were isolated, exhibiting variable degrees of PGP traits, including phosphate solubilization (10-375 µg/mL), indole-3-acetic acid (6-66 µg/mL), siderophore (32.37%-301.48% SU) production and antifungal activity in terms of percent growth inhibition (% GI) against Fusarium oxysporum (44.44%-77.77% GI), Fusarium graminearum (48.88%-71.42% GI) and Rhizoctonia solani (44.44%-77.7% GI). The 16S rDNA sequencing results showed lineage of these strains to 15 genera viz., Aneurinibacillus, Bacillus, Beijerinckia, Cedecea, Ensifer, Enterobacter, Kosakonia, Lactobacillus, Lysobacter, Oxynema, Pseudomonas, Pantoea, Paenibacillus, Pseudoxanthomonas and Serratia. Out of 42 strains, 11 potential strains were selected for in vivo growth studies of R. officinalis. The results showed that the inoculation of Bacillus subtilis KU21, Pseudomonas aeruginosa SI12, and Cedecea lapagei KU14 significantly increased the physical growth parameters of plant over uninoculated control viz., number of lateral of branches (43.95%-46.39%), stem height (29.04%-38.57%), root length (32.31%-37.14%), shoot (34.76%-40.91%) and root biomass (62.89%-70.70%). Physiological characteristics such as total chlorophyll (30.41%-30.96%), phenol (14.43%-24.55%) and carotenoids (34.26%-39.87%) content, also showed a relative increase as compared to uninoculated control; furthermore, the macronutrients (NPK) contents of the plant as well as soil also showed an increase. The developed module may be recommended for sustainable production of R. officinalis in the North-Western Himalayan region without hampering the soil health and fertility.

Transcriptome Analysis Reveals Differentially Expressed Genes That Regulate Biosynthesis of the Active Compounds with Methyl Jasmonate in Rosemary Suspension Cells.

To study the effects of Methyl jasmonates (MeJA) on rosemary suspension cells, the antioxidant enzymes' change of activities under different concentrations of MeJA, including 0 (CK), 10 (M10), 50 (M50) and 100 µM MeJA (M100). The results demonstrated that MeJA treatments increased the activities of phenylalanine ammonla-lyase (PAL), superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and polyphenol oxidase (PPO) and reduced the contents of hydrogen peroxide (H2O2) and malondialdehyde (MDA), thus accelerating the ROS scavenging. Comparative transcriptome analysis of different concentrations of MeJA showed that a total of 7836, 6797 and 8310 genes were differentially expressed in the comparisons of CKvsM10, CKvsM50, CKvsM100, respectively. The analysis of differentially expressed genes (DEGs) showed phenylpropanoid biosynthesis, vitamin B6, ascorbate and aldarate metabolism-related genes were significantly enriched. The transcripts of flavonoid and terpenoid metabolism pathways and plant hormone signal transduction, especially the jasmonic acid (JA) signal-related genes, were differentially expressed in CKvsM50 and CKvsM100 comparisons. In addition, the transcription factors (TFs), e.g., MYC2, DELLA, MYB111 played a key role in rosemary suspension cells under MeJA treatments. qRT-PCR of eleven DEGs showed a high correlation between the RNA-seq and the qRT-PCR result. Taken together, MeJA alleviated peroxidative damage of the rosemary suspension cells in a wide concentration range via concentration-dependent differential expression patterns. This study provided a transcriptome sequence resource responding to MeJA and a valuable resource for the genetic and genomic studies of the active compounds engineering in rosemary.

The effect of anaerobic digestate derived composts on the metabolite composition and thermal behaviour of rosemary.

The study reports on the effect of anaerobic digestate derived composts on the metabolite composition and thermal behaviour of rosemary (Rosmarinus officinalis L.). Plants were cultivated in semiarid soil under four different fertiliser treatments (composts of anaerobic digested cattle (C) or pig slurry (P) at 30t/ha and 60 t/ha, and two control treatments (inorganic fertiliser and no fertiliser application). Samples of leaves and stems were analysed to investigate the effect of treatment on chemical composition and thermochemical properties. Three orthogonal analytical approaches were used, namely: Fourier transform mid infrared spectroscopy (FTIR), gas chromatography/mass spectrometry (GC/MS) and thermochemical gravimetric analysis (TGA). FTIR and GC/MS showed fertiliser treatment resulted in tissue specific changes in sample metabolite composition. Fertiliser treatment was detected to change the thermogravimetric properties of the leaf samples and from inorganic and composted pig slurry digestate treatments had greater ash content and lower proportions of fixed carbon compared with samples from the unfertilised control treatment. This study provides information on how the composition of rosemary might be altered by fertiliser application in regions of poor soil, and has implications for biomass quality when rosemary is grown on semi-wild sites for the purpose of soil improvement.

Modified distribution in the polyphenolic profile of rosemary leaves induced by plant inoculation with an arbuscular mycorrhizal fungus.

BACKGROUND: Rosemary forms an arbuscular mycorrhizal (AM) symbiosis with a group of soilborne fungi belonging to the phylum Glomeromycota, which can modify the plant metabolome responsible for the antioxidant capacity and other health beneficial properties of rosemary. RESULTS: The effect of inoculating rosemary plants with an AM fungus on their growth via their polyphenolic fingerprinting was evaluated after analyzing leaf extracts from non-inoculated and inoculated rosemary plants by ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Plant growth parameters indicated that mycorrhizal inoculation significantly increased plant height and biomass. Chemical modifications in the plant polyphenolic profile distribution were found after a principal components analysis (PCA) loading plots study. Four compounds hosting strong antioxidant properties - ferulic acid, asiatic acid, carnosol, and vanillin - were related to mycorrhizal rosemary plants while caffeic and chlorogenic acids had a higher influence on non-mycorrhizal plants. CONCLUSION: Mycorrhization was found to stimulate growth to obtain a higher biomass of plant leaves in a short time, avoiding chemical fertilization, while analytical results demonstrate that there is an alteration in the distribution of polyphenols in plants colonized by the symbiotic fungus, which can be related to an improvement in nutritional properties with future industrial significance. © 2018 Society of Chemical Industry.

Constituents of Mediterranean Spices Counteracting Vascular Smooth Muscle Cell Proliferation: Identification and Characterization of Rosmarinic Acid Methyl Ester as a Novel Inhibitor.

SCOPE: Aberrant vascular smooth muscle cell (VSMC) proliferation is involved in atherosclerotic plaque formation and restenosis. Mediterranean spices have been reported to confer cardioprotection, but their direct influence on VSMCs has largely not been investigated. This study aims at examining rosmarinic acid (RA) and 11 related constituents for inhibition of VSMC proliferation in vitro, and at characterizing the most promising compound for their mode of action and influence on neointima formation in vivo. METHODS AND RESULTS: RA, rosmarinic acid methyl ester (RAME), and caffeic acid methyl ester inhibit VSMC proliferation in a resazurin conversion assay with IC50 s of 5.79, 3.12, and 6.78 µm, respectively. RAME significantly reduced neointima formation in vivo in a mouse femoral artery cuff model. Accordingly, RAME leads to an accumulation of VSMCs in the G0 /G1 cell-cycle phase, as indicated by blunted retinoblastoma protein phosphorylation upon mitogen stimulation and inhibition of cyclin-dependent kinase 2 in vitro. CONCLUSION: RAME represses PDGF-induced VSMC proliferation in vitro and reduces neointima formation in vivo. These results recommend RAME as an interesting compound with VSMC-inhibiting potential. Future metabolism and pharmacokinetics studies might help to further evaluate the potential relevance of RAME and other spice-derived polyphenolics for vasoprotection.

Report: Regional variation in the chemical composition and antioxidant activity of Rosmarinus officinalis L. from China and the Mediterranean region.

Rosemary is an aromatic evergreen shrubby herb that is native to the Mediterranean region. This herb is now widely cultivated in many regions of the world. Rosemary is widely used in traditional Chinese medicines, foods, nutraceuticals and cosmetics. Hydro distilled essential oils, obtained from rosemary in China and the Mediterranean region, were analysed by gas chromatography-mass spectrometry (GC-MS). Thirty-seven compounds accounting for 94.97%-99.72% of the oils were identified. The majority of the compounds in the essential oils exhibited no significant differences (table 1 and fig. 1). The extracts were prepared with three solvents of different polarity (dichloromethane, ethyl acetate and aqueous). The ethyl acetate fractions exhibited the highest phenol content and were found to be significantly more active than the dichloromethane and aqueous fractions (fig. 2). Antioxidant activity (by DPPH radical scavenging, ferric ion reduction (FRAP) and thiobarbituric acid reactive substance (TBARS)) was also assessed. The ethyl acetate extracts of Yunnan had the highest amount of antioxidant capacity from China by DPPH and TBARS, with the lowest IC50 values being 0.0011 mg/ml, and 1.6611 mg/ml, respectively. In conclusion, the antioxidant activities of the essential oils and ethyl acetate extracts from rosemary obtained by three different testing methods revealed higher antioxidant activity from rosemary grown in China than in the Mediterranean region. These results suggested that Chinese rosemary should be widely used in food, traditional medicine, cosmetics and perfume products, as well as other chemical industries.

Season-dependent and independent responses of Mediterranean scrub to light conditions.

Semi-arid plant species cope with excess of solar radiation with morphological and physiological adaptations that assure their survival when other abiotic stressors interact. At the leaf level, sun and shade plants may differ in the set of traits that regulate environmental stressors. Here, we evaluated if leaf-level physiological seasonal response of Mediterranean scrub species (Myrtus communis, Halimium halimifolium, Rosmarinus officinalis, and Cistus salvifolius) depended on light availability conditions. We aimed to determine which of these responses prevailed independently of the marked seasonality of Mediterranean climate, to define a leaf-level strategy in the scrub community. Thirty six leaf response variables - involving gas exchange, water status, photosystem II photochemical efficiency, photosynthetic pigments and leaf structure - were seasonally measured in sun exposed and shaded plants under field conditions. Physiological responses showed a common pattern throughout the year, in spite of the marked seasonality of the Mediterranean climate and of species-specific differences in the response to light intensity. Variables related to light use, CO2 assimilation, leaf pigment content, and LMA (leaf mass area) presented differences that were consistent throughout the year, although autumn was the season with greater contrast between sun and shade plants. Our data suggest that in Mediterranean scrub shade plants the lutein pool could have an important role in the photoprotection of the photosynthetic tissues. There was a negative linear correlation between the ratio lutein/total chlorophylls and the majority of leaf level variables. The combined effect of abiotic stress factors (light and drought or light and cold) was variable-specific, in some cases enhancing differences between sun and shade plants, while in others leading to unified strategies in all scrub species.

Differences in the composition of arbuscular mycorrhizal fungal communities promoted by different propagule forms from a Mediterranean shrubland.

As it is well known, arbuscular mycorrhizal (AM) colonization can be initiated from the following three types of fungal propagules: spores, extraradical mycelium (ERM), and mycorrhizal root fragments harboring intraradical fungal structures. It has been shown that biomass allocation of AM fungi (AMF) among these three propagule types varies between fungal taxa, as also differs the ability of the different AMF propagule fractions to initiate new colonizations. In this study, the composition of the AMF community in the roots of rosemary (Rosmarinus officinalis L., a characteristic Mediterranean shrub), inoculated with the three different propagule types, was analyzed. Accordingly, cuttings from this species were inoculated with either AMF spores, ERM, or colonized roots extracted from a natural soil. The AMF diversity within the rosemary roots was characterized using terminal restriction fragment length polymorphism (T-RFLP) of the small subunit (SSU) rDNA region. The AMF community established in the rosemary plants was significantly different according to the type of propagule used as inoculum. AMF taxa differed in their ability to initiate new colonizations from each propagule type. Results suggest different colonization strategies for the different AMF families involved, Glomeraceae and Claroideoglomeraceae colonizing mainly from colonized roots whereas Pacisporaceae and Diversisporaceae from spores and ERM. This supports that AMF taxa show contrasting life-history strategies in terms of their ability to initiate new colonizations from the different propagule types. Further research to fully understand the colonization and dispersal abilities of AMF is essential for their rational use in ecosystem restoration programs.

Phytostabilisation with Mediterranean shrubs and liming improved soil quality in a pot experiment with a pyrite mine soil.

Phytoremediation can be a suitable option to manage derelict mine soils. A pot experiment was carried out under semi-controlled conditions with a mine-impacted soil. A further contamination event was mimicked by applying 5% of pyritic sludge. Four species were planted in pots (Myrtus communis, Retama sphaerocarpa, Rosmarinus officinalis and Tamarix gallica), and some pots remained unplanted as a control. The substrates were moderately to highly contaminated, mainly with arsenic and zinc. The strong acidification induced by the pyritic sludge was buffered with lime and plants survived in all the pots. Liming provoked an effective immobilisation of metals and arsenic. Plant establishment decreased labile As in the substrate by 50%, mainly M. communis, although the levels of extractable metals were not affected by the plants. R. sphaerocarpa and M. communis increased the levels of C and N in the soil by 23% and 34% respectively, and also enhanced enzymatic activities and microbial respiration to the double in some cases. The low transfer of trace elements to shoots limited the phytoextraction rate. Our results support the use of phytostabilisation in Mediterranean mine soils and show how plants of R. sphaerocarpa and M. communis may increase soil health and quality during revegetation.

Phytoextraction of cadmium by four Mediterranean shrub species.

The possibility of remediating contaminated soils though the use of high biomass-generating, native plant species capable of removing heavy metals is receiving increased attention. The cadmium (Cd) accumulation capacities of the native Mediterranean, perennial shrubs Atriplex halimus, Phyllirea angustifolia, Rhamnus alaternus and Rosmarinus officinalis were tested by growing transplanted specimens in a pine bark compost substrate (pH 5.6) contaminated with 100 mg Cd kg(-1). After 70 days, only R. alaternus showed reduced growth. The increase in biomass seen in all the test species enhanced the phytoextraction of Cd. However, the species behaved as metal excluders, except for the halophyte A. halimus, which behaved as an indicator plant. In this species the leaf Cd concentration reached 35 mg Cd kg(-1), with the shoot responsible for some 86% of total Cd accumulation. Atriplex halimus showed the highest bioconcentration factor (BCF) (0.36) and leaf Cd transport index (1.68); consequently, this species showed the highest Cd phytoextraction capacity.

Fenologia do alecrim-pimenta (Lippia sidoides Cham. ) em área de Cerrado, no norte de Minas Gerais, Brasil / Pepper-rosmarin (Lippia sidoides Cham. ) phenology in a Cerrado area in northern Minas Gerais State, Brazil

Este trabalho teve como objetivo descrever o comportamento fenológico de Lippia sidoides Cham., em área de Cerrado, no norte de Minas Gerais, buscando-se subsidiar o processo de domesticação da espécie, por meio do estudo da ocorrência e duração das fenofases vegetativas e reprodutivas da espécie, bem como avaliar a influência dos fatores climáticos da região em cada fenofase, e ainda verificar a sincronia dessas fenofases entre os indivíduos. Vinte indivíduos foram avaliados mensalmente no período de fevereiro de 2007 a fevereiro de 2009, quanto à intensidade e sincronia das fenofases de brotamento, folhas maduras, senescência foliar, floração, frutificação e queda foliar. Houve acentuada periodicidade na ocorrência dos eventos fenológicos foliares, sendo que essa periodicidade acompanhou a estacionalidade climática da região, evidenciando a forte influência dos fatores abióticos, principalmente da precipitação e umidade relativa do ar. O alecrim-pimenta apresentou comportamento caducifólio, com alta taxa de queda foliar durante grande parte do ano. A floração ocorreu no período de fevereiro a abril, enquanto a frutificação alcançou o pico de intensidade no mês de maio. Todas as fenofases apresentaram altos índices de sincronia, sendo esse um fator facilitador do manejo da espécie.

The aim of this work was to describe the phenological behavior of Lippia sidoides Cham. in a Cerrado area in northern Minas Gerais State, Brazil, in order to contribute to the process of domestication of this species by studying the occurrence and duration of its vegetative and reproductive phenophases, evaluating the influence of local climatic factors on each phenophase and investigating the synchrony of these phenophases among individuals. Twenty specimens were monthly evaluated between February 2007 and February 2009 as to intensity and synchrony of the phenophases sprouting, mature leaves, leaf senescence, flowering, fruiting and leaf abscission. There was great periodicity in the occurrence of leaf phenological events according to the local climatic seasonality, highlighting the strong influence of abiotic factors, especially rainfall and relative air humidity. Pepper-rosmarin showed deciduous behavior, with high leaf abscission rate during a great part of the year. Flowering occurred from February to April, whereas fruiting peaked in May. All phenophases had high levels of synchrony, a facilitating factor for the management of this species.

Produção de fitomassa e teor de óleo essencial de folhas de alecrim-pimenta (Lippia sidoides Cham. ) em diferentes espaçamentos de plantio / Phytomass production and essential oil yield of pepper-rosmarin (Lippia sidoides Cham. ) leaves in different planting spacings

O alecrim-pimenta, espécie nativa do nordeste brasileiro, é uma planta medicinal de porte arbustivo, com folhas aromáticas, que possuem óleo essencial rico em timol e carvacrol. Essas substâncias conferem grande importância à planta, uma vez que apresentam atividades farmacológicas comprovadas cientificamente. Desta forma, o estudo das condições de cultivo da espécie é essencial para obtenção de maiores produtividades. O espaçamento entre plantas destaca-se como um dos fatores mais importantes relativos ao cultivo, de forma que a produtividade e o rendimento da cultura são afetados pela densidade de plantas. Dada a necessidade de otimizar técnicas de cultivo do alecrim-pimenta (Lippia sidoides Cham.), objetivou-se avaliar a produção de fitomassa e óleo essencial em quatro espaçamentos de plantio. O trabalho foi realizado em campo, no Instituto de Ciências Agrárias da Universidade Federal de Minas Gerais (ICA/UFMG), no município de Montes Claros/MG. Utilizou-se o delineamento em blocos casualizados, com quatro tratamentos e cinco repetições. Os tratamentos foram definidos pelos espaçamentos de plantio: 1,0 x 0,5 m; 1,0 x 1,0 m; 1,5 x 1,0 m; e fileiras duplas (1,0 x 0,8 x 0,5m), sendo 1,0 m entre fileiras duplas, 0,8 m entre fileiras simples e 0,5 m entre plantas na fileira. Após 150 dias do plantio no campo, foram avaliadas as variáveis produção de fitomassa fresca e seca das folhas, a produção e o teor de óleo essencial. Observou-se que o espaçamento de 1,0 x 0,5 m possibilitou uma maior produção de fitomassa e de óleo essencial, sendo, portanto, o mais indicado para o cultivo do alecrim-pimenta (Lippia sidoides Cham.).

Native to Northeastern Brazil, pepper-rosmarin (Lippia sidoides Cham.) is a shrubby medicinal plant with aromatic leaves and essential oil rich in thymol and carvacrol. These components have great importance to the species since they present scientifically proven pharmacological activities. Thus, the study of cultivation conditions for this species is essential to obtain higher productivity. Spacing between plants is one of the most important factors related to cultivation, since crop productivity and yield are affected by plant density. Based on the need of optimizing pepper-rosmarin cultivation techniques, this work aimed to evaluate the phytomass production and the essential oil yield in four planting spacings. The experiment was carried out under field conditions in the Agricultural Sciences Institute, Federal University of Minas Gerais (ICA/UFMG), Montes Claros Municipality, Minas Gerais State, Brazil. Experimental design was in randomized blocks, with four treatments and five replicates. Treatments consisted of the planting spacings 1.0 x 0.5 m; 1.0 x 1.0 m; 1.5 x 1.0 m; and double rows (1.0 x 0.8 x 0.5 m), with 1.0m between double rows, 0.8m between single rows and 0.5m between plants in the row. After 150 days of planting in the field, leaf fresh and dry phytomass production and essential oil yield were evaluated. The spacing 1.0 x 0.5 m led to higher phytomass production and essential oil yield, being therefore the most recommended for pepper-rosmarin cultivation.

Produção de mudas de dois genótipos de alecrim-de-tabuleiro (Lippia gracilis Schauer) em função de fertilizante mineral, calcário, substratos e recipientes / Seedling production of two "alecrim-de-tabuleiro" (Lippia gracilis Schauer) genotypes under mineral fertilizer, limestone, substrates and containers

Lippia gracilis Schauer popularmente conhecida como alecrim-de-tabuleiro possui moderada atividade antibacteriana, antimicrobiana e antiséptica. Objetivando avaliar o efeito de doses de fertilizante mineral, calcário, substratos e recipientes na produção de mudas de dois genótipos de alecrim-de-tabuleiro realizou-se experimentos com estacas de dois genótipos de L. gracilis. No experimento 1, foram utilizadas estacas apicais distribuídas em bandejas de poliestireno expandido de 128 células com o substrato pó-de-coco + areia (1:1), três repetições e oito estacas por repetição. O delineamento experimental foi em blocos ao acaso, em esquema fatorial 4x2x2, sendo quatro doses do fertilizante (6-24-12+micronutrientes) (1, 2, 3 e 4 g L-1), duas doses de calcário (0 e 1 g L-1) e dois genótipos (LGRA106 e LGRA201). Aos 35 após plantio, foram avaliadas a sobrevivência ( por cento), enraizamento ( por cento), comprimento de raiz (cm) e massa seca de raiz (mg). No experimento 2, foram utilizadas três repetições com oito estacas por repetição. O delineamento experimental foi em blocos ao acaso, em esquema de parcelas subdivididas onde foram testados nas parcelas dois recipientes (bandeja de poliestireno expandido de 128 alvéolos e tubetes de 110 cm³, na sub parcela foram testadas as combinações de quatro substratos (PC - pó de coco; PCA (1:1) - pó de coco + areia (1:1); PCA (2:1) - pó de coco + areia (2:1) e PCA (3:1) - pó de coco + areia (3:1) e dois genótipos (LGRA106 e LGRA201). Aos 35 dias após o plantio foram analisadas as mesmas variáveis do experimento 1, além de altura de parte aérea (cm) e massa seca de parte aérea (mg). A utilização de 1 g L-1 do fertilizante na ausência de calcário foi efetivo para sobrevivência de plantas e enraizamento de estacas de alecrim-de-tabuleiro. Com base nesses experimentos, concluiu-se que o genótipo LGRA106 é superior ao LGRA201 nas variáveis analisadas e recomenda-se a bandeja de poliestireno expandido para produção de mudas de L. gracilis via estaquia.

Commonly known as "alecrim-de-tabuleiro", Lippia gracilis Schauer (Verbenaceae) has moderate antibacterial, antimicrobial and antiseptic activity. This study aimed to evaluate the effect of doses of mineral fertilizer, limestone, substrates and containers on seedling production of two "alecrim-de-tabuleiro" genotypes, using cuttings. In experiment 1, apical cuttings were distributed on expanded polystyrene trays of 128 cells containing coir + sand (1:1), using three replicates of eight cuttings each. Experimental design was in randomized blocks, in a 4x2x2 factorial arrangement, i.e. four doses of fertilizer (6-24-12 + micronutrients) (1, 2, 3 and 4 g L-1), two doses of limestone (0 and 1 g L-1) and two genotypes (LGRA106 and LGRA201). At 35 days after planting, we evaluated survival ( percent), rooting ( percent), root length (cm) and root dry matter (mg). In experiment 2, three replicates of eight cuttings each were used. Experimental design was in randomized blocks, in a split-plot scheme. In the plots, we tested two containers (expanded polystyrene tray of 128 cells and 110 cm³ tubets). In the subplots, we tested four substrate combinations [coir, coir + sand (1:1), coir + sand (2:1) and coir + sand (3:1)] and two genotypes (LGRA106 and LGRA201). At 35 days after planting, the same variables of experiment 1 were evaluated, besides shoot length (cm) and shoot dry matter (mg). The use of 1 g L-1 fertilizer without limestone was effective for plant survival and cutting rooting. Based on these experiments, the genotype LGRA106 is superior to LGRA201 as to the evaluated variables and the expanded polystyrene tray is recommended for the production of L. gracilis seedlings through cuttings.

Produção de biomassa e óleo essencial de alecrim (Rosmarinus officinalis L. ) em função da altura e intervalo entre cortes / Biomass production and essential oil of rosemary (Rosmarinus officinalis L. ) in function of the height and interval between the cuts

O alecrim é planta exótica e o interesse no seu cultivo tem crescido ao longo dos anos, pois pode ser utilizado tanto para fins medicinais como aromáticos; no entanto, o conhecimento fitotécnico sobre a espécie é bastante limitado. Dessa forma, o trabalho objetivou avaliar a influência da altura de corte e do intervalo destes sobre a produção de biomassa e no rendimento do óleo essencial de alecrim. O experimento foi realizado no Instituto Agronômico, em Campinas - SP, no período de janeiro de 2006 a julho de 2007. O delineamento experimental utilizado foi em blocos ao acaso, com 8 tratamentos em esquema fatorial 2 x 4, sendo duas alturas de corte (20 e 40 cm) e quatro intervalos entre cortes (60, 80, 100, 120 dias) e três repetições. Foram avaliadas as características altura da planta, massa seca da parte aérea, rendimento e qualidade do óleo essencial. Observou-se que maiores intervalos entre cortes proporcionaram maior altura da planta e maior massa seca da parte aérea. O rendimento e a qualidade do óleo essencial não foram afetados ao longo do período experimental.

Rosemary is an exotic plant and the interest in its cultivation has been growing along the years, because it can be used for medicinal or aromatic ends, however the agricultural knowledge about the species is quite limited. The goal of this research was to evaluate the influence of the cutting height and of the interval of these cuts on the biomass production and on the yield of the rosemary essential oil. The experiment was conducted at the Agronomical Institute, in Campinas - SP, from January 2006 through July 2007. It was organized in a completely randomized design with 8 treatments in a 2 x 4 factorial system, with two cutting heights (20 and 40 cm) and four intervals between cuts (60, 80, 100, 120 days) and three replications. Characteristics such as plant height, dry mass of the aerial part, yield and quality of its essential oil were evaluated. It was observed that longer intervals among cuttings provided longer height of the plant and greater dried mass of the aerial part. The yield and the quality of the essential oil were not affected throughout the experimental period.

Production and diversity of volatile terpenes from plants on calcareous and siliceous soils: effect of soil nutrients.

Fertilizer effects on terpene production have been noted in numerous reports. In contrast, only a few studies have studied the response of leaf terpene content to naturally different soil fertility levels. Terpene content, as determined by gas chromatography/mass spectrometry/flame ionization detector, and growth of Pinus halepensis, Rosmarinus officinalis, and Cistus albidus were studied on calcareous and siliceous soils under field conditions. The effect of nitrogen (N) and extractable phosphorus (P(E)) from these soils on terpenes was also investigated since calcareous soils mainly differ from siliceous soils in their higher nutrient loadings. Rich terpene mixtures were detected. Twenty-one terpenes appeared in leaf extracts of R. officinalis and C. albidus and 20 in P. halepensis. Growth of all species was enhanced on calcareous soils, while terpene content showed a species-specific response to soil type. The total monoterpene content of P. halepensis and that of some major compounds (e.g., delta-terpinene) were higher on calcareous than on siliceous soils. A significant and positive relationship was found between concentration of N and P(E) and leaf terpene content of this species. These findings suggest that P. halepensis may respond to an environment characterized by increasing soil deposition, by allocating carbon resources to the synthesis of terpene defense metabolites without growth reduction. Results obtained for R. officinalis showed high concentrations of numerous major monoterpenes (e.g., myrcene, camphor) in plants growing on calcareous soils, while alpha-pinene, beta-caryophyllene, and the total sesquiterpene content were higher on siliceous soils. Finally, only alloaromadendrene and delta-cadinene of C. albidus showed higher concentrations on siliceous soils. Unlike P. halepensis, soil nutrients were not involved in terpene variation in calcareous and siliceous soils of these two shrub species. Possible ecological explanations on the effect of soil type for these latter two species as well as the ecological explanation of rich terpene mixtures are discussed.

Variation of the chemical profile and antioxidant behavior of Rosmarinus officinalis L. and Salvia fruticosa Miller grown in Greece.

In this study, the essential oil and the phenolic composition along with the antioxidant activity of R. officinalis L. and S. fruticosa Miller, collected in Zakynthos island (Ionian Sea, Greece), were investigated. The essential oil composition of the plants was characterized by the presence of 1,8-cineole. Mean values of the antioxidant activities of rosemary and sage essential oils indicated slight differences. The antioxidant activity of sage oil was correlated with the oxygenated sesquiterpenes and diterpenes concentrations. Concerning the methanolic extracts, a close relationship between the phenolic content and the development stage during vegetative cycle of these plants was observed. The identified flavonoids, except rutin, seemed to increase with the advancement of developmental stages, while phenolic acids followed an opposite pattern. The antioxidant activity was correlated with the amount of total phenolic content.

Enhanced carnosic acid levels in two rosemary accessions exposed to cold stress conditions.

Two rosemary accessions were subjected to chilling temperatures in control environmental cabins analyzing their variations in rosmarinic and carnosic acids together with their adaptability to these stress conditions. Cold stressed plants of both accessions showed increases in caffeic acid and carnosic acid concentration levels, while other secondary metabolites such as rosmarinic acid, naringin, cirsimaritin, hispidulin, and carnosol showed different responses in both accessions. In addition, cold stressed plants exhibited significant reductions in chlorophylls, beta-carotene, and violaxanthin levels as well as the maximum quantum yield of PSII in both accessions. Hydrogen peroxide and lipid peroxidation levels showed similar responses in both accessions, which were positively and negatively correlated with rosmarinic and carnosic acids. From these results it is therefore suggested that carnosic acid biosynthesis in rosemary plants is induced by chilling periods. On the other hand, we demonstrate that not all rosemary accessions are equally well adapted to chilling temperatures. In fact, for (one) accession cold treated plants severe losses in chlorophyll, beta-carotene, and even xanthophylls (including zeaxanthin and antheraxanthin) were observed, despite no visual symptoms of leaf injury. More research is needed to understand rosmarinic acid variations in rosemary plants under stress conditions.

Effects of irrigation and air humidity preconditioning on water relations, growth and survival of Rosmarinus officinalis plants during and after transplanting.

The effect of different irrigation and air humidity conditioning treatments on the morphological and physiological responses of Rosmarinus officinalis in nursery conditions was investigated in order to evaluate the degree of hardening resulting from these conditions. Rosmarinus officinalis seedlings were pot-grown during 4 months in two greenhouses (nursery period), in which two irrigation treatments were used (control and deficit). In one of these greenhouses, air humidity was controlled using a dehumidifying system (low humidity), in the other greenhouse the air conditions were not artificially modified (control humidity). After the nursery period, the plants of all treatments were transplanted and well watered (100% water holding capacity for 1 month, transplanting period). After this period, they received no water (establishment period). At the end of the nursery period it was seen that deficit irrigation had altered the morphology of the R. officinalis plants by reducing plant height, stem diameter, leaf area, total dry weight, and root length, while humidity influenced the parameters related with plant water relations. Low air humidity and deficit irrigation-induced tissue dehydration and lower stomatal conductance values (gs). The plants subjected to deficit irrigation developed leaf osmotic adjustment, which was maintained during the transplanting period. At that time, the plants that had been exposed to deficit irrigation and low humidity showed efficient stomatal regulation (lower gs values). After transplanting and during the establishment period, these plants showed a better water status (higher psil and gs values). Their post-planting survival rate improved as a result of acclimation processes.

Flavonoid distribution during the development of leaves, flowers, stems, and roots of Rosmarinus officinalis. postulation of a biosynthetic pathway.

The distribution of seven flavonoids, eriocitrin, luteolin 3'-O-beta-d-glucuronide, hesperidin, diosmin, isoscutellarein 7-O-glucoside, hispidulin 7-O-glucoside, and genkwanin, has been studied in Rosmarinus officinalis leaves, flowers, stems, and roots during plant growth. The maximum level reached by luteolin 3'-O-beta-d-glucuronide in leaves during June-August suggests the existence of a delay between the activation of the enzymes involved in the flavanone and flavone biosynthesis. The presence of hesperidin and diosmin in the vascular system is significant, and hesperidin shows even higher levels than the phenolic diterpenes and rosmarinic acid. The distribution of flavonoids observed in R. officinalis suggests a functional and structural relationship between phytoregulators and flavonoids, where flavonoids would be "protectors" of the activity of phytoregulators. A hypothesis for the general pathway of biosynthesis of these compounds in plants of the family Labiatae is proposed.

Variations in water status, gas exchange, and growth in Rosmarinus officinalis plants infected with Glomus deserticola under drought conditions.

The influence of the arbuscular mycorrhizal fungus Glomus deserticola on the water relations, gas exchange parameters, and vegetative growth of Rosmarinus officinalis plants under water stress was studied. Plants were grown with and without the mycorrhizal fungus under glasshouse conditions and subjected to water stress by withholding irrigation water for 14 days. Along the experimental period, a significant effect of the fungus on the plant growth was observed, and under water stress, mycorrhizal plants showed an increase in aerial and root biomass compared to non-mycorrhizal plants. The decrease in the soil water potential generated a decrease in leaf water potential (psi(l)) and stem water potential (psi(x)) of mycorrhizal and non-mycorrhizal plants, with this decrease being lower in mycorrhizal water-stressed plants. Mycorrhization also had positive effects on the root hydraulic conductivity (Lp) of water stressed plants. Furthermore, mycorrhizal-stressed plants showed a more important decrease in osmotic potential at full turgor (psi(os)) than did non-mycorrhizal-stressed plants, indicating the capacity of osmotic adjustment. Mycorrhizal infection also improved photosynthetic activity (Pn) and stomatal conductance (g(s)) in plants under water stress compared to the non-mycorrhizal-stressed plants. A similar behaviour was observed in the photochemical efficiency of PSII (Fv/Fm) with this parameter being lower in non-mycorrhizal plants than in mycorrhizal plants under water stress conditions. In the same way, under water restriction, mycorrhizal plants showed higher values of chlorophyll content than did non-mycorrhizal plants. Thus, the results obtained indicated that the mycorrhizal symbiosis had a beneficial effect on the water status and growth of Rosmarinus officinalis plants under water-stress conditions.