Transcriptome-Wide Identification and Characterization of the JAZ Gene Family in Mentha canadensis L.

Jasmonate ZIM-domain (JAZ) proteins are the crucial transcriptional repressors in the jasmonic acid (JA) signaling process, and they play pervasive roles in plant development, defense, and plant specialized metabolism. Although numerous JAZ gene families have been discovered across several plants, our knowledge about the JAZ gene family remains limited in the economically and medicinally important Chinese herb Mentha canadensis L. Here, seven non-redundant JAZ genes named McJAZ1-McJAZ7 were identified from our reported M. canadensis transcriptome data. Structural, amino acid composition, and phylogenetic analysis showed that seven McJAZ proteins contained the typical zinc-finger inflorescence meristem (ZIM) domain and JA-associated (Jas) domain as conserved as those in other plants, and they were clustered into four groups (A-D) and distributed into five subgroups (A1, A2, B1, B2, and D). Quantitative real-time PCR (qRT-PCR) analysis showed that seven McJAZ genes displayed differential expression patterns in M. canadensis tissues, and preferentially expressed in flowers. Furthermore, the McJAZ genes expression was differentially induced after Methyl jasmonate (MeJA) treatment, and their transcripts were variable and up- or down-regulated under abscisic acid (ABA), drought, and salt treatments. Subcellular localization analysis revealed that McJAZ proteins are localized in the nucleus or cytoplasm. Yeast two-hybrid (Y2H) assays demonstrated that McJAZ1-5 interacted with McCOI1a, a homolog of Arabidopsis JA receptor AtCOI1, in a coronatine-dependent manner, and most of McJAZ proteins could also form homo- or heterodimers. This present study provides valuable basis for functional analysis and exploitation of the potential candidate McJAZ genes for developing efficient strategies for genetic improvement of M. canadensis.

How the morphological properties of Mentha longifolia.(L.) Huds. may be affected by geographical differences.

With respect to the benefits of medicinal plants for human health and their use as food, they are extensively being researched and planted in different parts of the world including Iran. Among the most important parameters, which may affect the growth and the yield of medicinal plants is their geographical (ecotype) properties. Accordingly, a research work was proposed to investigate the effects of 10 different regions or ecotypes on the morphological properties of pennyroyal [Mentha longifolia. (L.) Huds.] from the Lamiaceae family in one of the Iranian provinces, namely Ilam. According to the geographical characteristics of the experimental province, the longitudes were in the range of 60-65°, the latitudes varied from 36-37° and the heights above the sea level were in the range of 860-1410m. The experiment was a completely randomized block design in three replicates and for each plant ecotype, 20 plant samples in three replicates were collected. Different plant morphological properties related to plant stem, leaf, inflorescence, rhizome, and dry weight were determined. Data were subjected to analysis of variance and different plant statistical properties were determined. Using the Ward's method, the cluster analysis of data was also conducted and similar ecotypes were grouped. Data indicated the significant effects of regional (ecotype) differences on plant morphological properties and dry weight. The highest stem length was related to the Alamdar region (90cm), stem diameter and leaf number to Kolm region at 0.76cm and 128, respectively, leaf length to Chenarbashi at 4.07cm, leaf width to Helaesm at 2.83cm, inflorescence number and length to Sarab at 22.10 and 11.63cm, respectively, inflorescence width to Kolm at 1.04cm, rhizome length to Mishkhas at 11.00cm, and rhizome width to Kolm at 1.03cm. The highest dry weights were resulted by Siahkal (260g), Kolm (205g) and Chemarbashi (220g). Interestingly, stem length (22.17), inflorescence width (12.12) and rhizome length (27.11) indicated the least rate of coefficient of variation and had the highest effects on plant growth and yield production. Data indicated how geographical and soil properties (such as organic matter) may affect pennyroyal growth and yield. Such analyses can be used for the selection and the subsequent domestication of wild populations of pennyroyal with the highest efficiency.

Physiological, biochemical and molecular responses of Mentha aquatica L. to manganese.

Mentha aquatica is an aromatic herb which possesses valuable terpenoids constituents. Here, we intended to evaluate the effects of the different manganese (Mn) concentrations on the physiological, biochemical and molecular responses in M. aquatica. Basic Hoagland's solution (control), 40, 80, and 160 µM of Mn supplied as MnSO4·H2O were applied to the nutrient solution. The results indicated that the different concentrations of Mn differently affected the physiological, biochemical and molecular responses in M. aquatica. The growth parameters (biomass and photosynthetic pigments) and expression levels of ß-caryophyllene synthase (CPS), limonene synthase (Ls), geranyl diphosphate synthase (Gpps), and menthofuran synthase (Mfs) genes were increased at the moderate Mn concentrations (40 and 80 µM) and began to decrease at the higher levels. However, the contents of anthocyanins, flavonoids, malonaldehyde (MDA) and hydrogen peroxide (H2O2), Mn accumulation, activities of antioxidant enzymes, yield of essential oils and the expression levels of 1-Deoxy d-xylulose-5-phosphate synthase (Dxs) and isopentenyl diphosphate isomerase (Ippi) genes were gradually increased with increasing concentration of Mn in the nutrient solution. Also, the content and chemical composition of terpenoid constituents were altered in the Mn-treated plants. Here, we suggest that the application of external Mn in nutrient solution elevates the growth and expression levels of the genes that are involved in the terpenoid biosynthesis pathway in M. aquatica. Nevertheless, the extent and stability of these growth and gene expression elevation are varied among the different Mn treatments.

[Habitat suitability and quality division of Mentha haplocalyx].

In this study, ecological factors, occurrence records, the essential oil components content were used to predict the potential geographical distribution and quality division of Mentha haplocalyx in China based on the MaxEnt modeling and geographic information system(GIS). The AUC of ROC curve was above 0.950,indicating that the predictive results with the maximum model were highly precise. The results showed that the main environmental factors determining the potential distribution were annual average precipitation (the contribution rate, 45.87%), mean temperature of wettest quarter (11.92%), mean temperature of warmest quarter (7.84%), average monthly precipitation of May (6.80%), standard deviation of seasonal temperature variation (4.42%), mean temperature of the coldest quarter (3.47%) and altitude (2.92%). The environmental variables in the highly potential areas were determined as annual average precipitation around [530,1 465] mm, mean temperature of wettest quarter around [24.5,29] ℃, mean temperature of the warmest quarter around [25.5,29] ℃, average monthly precipitation of May around [67,133] mm, standard deviation of temperature seasonal change around [8 333,9 643], mean temperature of the coldest quarter around [1.7,8.3] ℃ and the altitude around [0,165] mm. The best quality distribution of M. haplocalyx was mainly located in Jiangsu, Anhui, Shandong, Zhejiang and Heilongjiang. The zoning results basically coincide with the actual situation. The quality division of M. haplocalyx can be used for providing a scientific basis for selection of artificial planting base and guidance of its production.

Impact assessment of mercury accumulation and biochemical and molecular response of Mentha arvensis: a potential hyperaccumulator plant.

The present study was focused on examining the effect of Hg oxidative stress induced physiochemical and genetic changes in M. arvensis seedlings. The growth rate of Hg treated seedlings was decreased to 56.1% and 41.5% in roots and shoots, respectively, compared to the control. Accumulation of Hg level in both roots and shoots was increased with increasing the concentration of Hg. Superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activities were found to be increased with increasing the Hg concentration up to 20 mg/L; however, it was decreased at 25 mg/L Hg concentration. The POX enzyme activity was positively correlated with Hg dose. The changes occurring in the random amplification of ploymorphic DNA (RAPD) profiles generated from Hg treated seedlings included variations in band intensity, disappearance of bands, and appearance of new bands compared with the control seedlings. It was concluded that DNA polymorphisms observed with RAPD profile could be used as molecular marker for the evaluation of heavy metal induced genotoxic effects in plant species. The present results strongly suggested that Mentha arvensis could be used as a potential phytoremediator plant in mercury polluted environment.

Performance of a wall cascade constructed wetland treating surfactant-polluted water.

Carwashes are highly water-consuming processes that require wastewater treatment before discharge into a sewer system due to the complex composition of leachate. Anionic surfactants (AS) are the main constituents of this wastewater because of their cleaning and solubilization properties; they can be potentially dangerous for the environment if not adequately treated. Constructed wetlands (CWs) are low-cost systems increasingly used to treat different types of wastewater; however, there are few studies on their use for the treatment of carwash wastewater. In this study, an innovative constructed wetland arranged in a "cascade" to simulate a wall system (WCCW) was experimented in 2010 and 2011 to treat AS. Three plant species were tested at different AS inlet concentrations (10, 50, and 100 mg L(-1)) with two hydraulic retention times (HRTs; 3 and 6 days): ribbon grass (Typhoides arundinacea (L.) Moench (syn. Phalaris arundinacea L.) var. picta; Ta), water mint (Mentha aquatica L.; Ma), and divided sedge (Carex divisa Hudson; Cd). All plant species grew constantly over the experimental period, showing a capacity to tolerate even the highest AS concentration. Using the HRT of 6 days, raising the inlet concentration increased the AS outlet concentration, with similar values for the treatments (median values of 0.13-0.15, 0.47-0.78, and 1.19-1.46 mg L(-1) at inlet concentrations in the order 10, 50, and 100 mg L(-1)). The shorter HRT led to significant differences among treatments in the reduction of outlet concentration, the best result being given by the tanks vegetated with Ma (A = 97.7 % with outlet concentration 0.35 mg L(-1)). After treatments of the WCCW, the AS content was reduced almost completely, with removal in the ranges 0.07-10.2 g m(-2) day(-1) for tanks planted with Ta, 0.10-9.1 g m(-2) day(-1) for Ma tanks, and 0.11-9.5 g m(-2) day(-1) for Cd tanks depending on the inlet concentration.

Organic C dynamics and its conservation under wheat (Triticum aesetivum) - mint (Mentha arvensis)-Sesbania rostrata cropping in sub-tropical condition of northern Indo-Gangetic plains.

Soil organic carbon (SOC) is accumulated or depleted as a result of cropping and management strategies. It plays a significant role in maintaining soil quality, plant productivity and mitigating greenhouse gas emission. We studied the long-term (20 years) influence of a wheat-mint-Sesbania cropping system on the SOC stock. Estimates of stabilization of SOC in different pools and a tentative C budget were also developed. Twenty years of cultivation caused a decrease in SOC only in control soils, which received no manure and fertilizer. However, it increased with balanced use of NPK inputs. Soil C stock decreased significantly with increased in soil depth 0-15 cm to 15-30 and 30-45 cm. About 6% (-2 to+14) of the C added in crop residues and green manure were stabilized in the soil. On an average 12%, 14%, 59%, 15% of the water stable aggregates were in the >2 mm, 2.0-0.25 mm, 0.25-0.05 mm, and <0-0.5 size fractions, respectively. Significant improvements in structural stability and nitrogen availability were detected in all the treatments compared to the control. The amount of organic C oxidizable by a modified Walkley and Black method, which involves using only half of the amount of sulphuric acid, is a more sensitive indicator of the improvement in soil quality parameters under investigation, namely SOC, and increases in mineralizable N and water stable aggregation than the standard method.

[Correlation between the output and composition of essential oil and the level of salicylic acid in mint plants at different ontogenetic stages].

Dynamic changes in the content of acetylsalicylic acid and the output and qualitative composition of essential oil have been studied in mint plants (Mentha spicata L. and cultivar Medichka) during their ontogenesis with allowance for changes in weather conditions. Ontogenetic changes in the level of acetylsalicylic acid in leaf tissues are found to be similar in both cv. Medichka and M. spicata. In the case of cv. Medichka, this change is connected with the dynamics of the production and the qualitative composition of essential oil; in the case of M. spicata, this connection is less expressed. The role of acetylsalicylic acid and essential oil in plant adaptation to the environment is discussed.

Effect of gibberellic acid and calliterpenone on plant growth attributes, trichomes, essential oil biosynthesis and pathway gene expression in differential manner in Mentha arvensis L.

Extensive research is going on throughout the world to find out new molecules from natural sources to be used as plant growth promoter. Mentha arvensis L. is the main source of menthol rich essential oil used commercially in various food, pharmaceutical and other preparations. Experiments were conducted on field grown plants for understanding the effect of calliterpenone (CA), a stereo-isomer of abbeokutone, in comparison to gibberellic acid (GA3) on growth attributes, trichomes, essential oil biosynthesis and expression of some oil biosynthetic pathway genes. The exogenous application of CA (1 µM, 10 µM and 100 µM) was found to be better in improving plant biomass and stolon yield, leaf area, branching and leaf stem ratio than with counterpart GA3 at the same concentrations. CA treated plants showed higher glandular trichome number, density and diameter and also correlated with enhanced oil biogenetic capacity as revealed by feeding labeled (14)C-sucrose for 72 h to excised shoots. Semi-quantitative PCR analysis of key pathway genes revealed differential up regulation under CA treatments. Transcript level of menthol dehydrogenase/menthone reductase was found highly up regulated in CA treated plants with increased content of menthone and menthol in oil. These findings demonstrate that CA positively regulated the yields by enhanced branching and higher density of trichomes resulting into higher accumulation of essential oil. The results suggest CA as a novel plant derived diterpenoid with growth promoting action and opens up new possibilities for improving the crop yields and essential oil biosynthesis in qualitative and quantitative manner.

The greater effectiveness of Glomus mosseae and Glomus intraradices in improving productivity, oil content and tolerance of salt-stressed menthol mint (Mentha arvensis).

BACKGROUND: Mentha arvensis is cultivated in large parts of the world for its menthol-rich essential oil. The study investigates the potential of four mycorrhizal fungi, viz. Glomus mosseae (Gm), Glomus aggregatum (Ga), Glomus fasciculatum (Gf) and Glomus intraradices (Gi) in alleviating NaCl-induced salt stress in Mentha arvensis cv. Kosi and establishes the specificity of interaction between different mycorrhizal species and their effectiveness in mitigating salt stress in Mentha arvensis. Mycorrhizal and non-mycorrhizal Mentha plants were subjected to NaCl-induced salinity. RESULTS: Among the four Glomus species, Gm and Gi reduced salt-induced herb yield losses: a loss of 27.53% and 25.58% respectively under salt stress in comparison to 51.00% in non-mycorrhizal M. arvensis salt-stressed plants. Gm- and Gi-inoculated plants also recorded higher leaf:stem ratio, oil content, and oil yield and menthol concentration in essential oil under both saline and non-saline conditions. CONCLUSION: Better performance in terms of herb yield, and oil content and yield was observed in Gi- and Gm-inoculated M. arvensis plants, suggesting the capability of Gi and Gm in protecting plants from the detrimental effects of salt stress; beneficial effects of arbuscular mycorrhizal fungi, however, may vary with host and environment.

A natural plant growth promoter calliterpenone from a plant Callicarpa macrophylla Vahl improves the plant growth promoting effects of plant growth promoting rhizobacteria (PGPRs).

Experiments were conducted to evaluate the efficacy of calliterpenone, a natural plant growth promoter from a shrub Callicarpa macrophylla Vahl., in enhancing the growth and yield promoting effects of plant growth promoting rhizobacteria (PGPRs), in menthol mint (Mentha arvensis L).This study is based on our previous results indicating the microbial growth promotion by calliterpenone and assumption that application of calliterpenone along with PGPRs will improve the population of PGPRs resulting in higher impacts on plant growth and yield. Of the 15 PGPRs (identified as potent ones in our laboratory), 25 µl of 0.01 mM calliterpenone (8.0 µg/100 ml) was found to be useful in improving the population of nine PGPRs in culture media. The five selected strains of PGPRs exhibiting synergy with calliterpenone in enhancing growth of maize compared to PGPR or calliterpenone alone were selected and tested on two cultivars (cvs. Kosi and Kushal) of M. arvensis. Of the five strains, Bacillus subtilis P-20 (16S rDNA sequence homologous to Accession No NR027552) and B. subtilis Daz-26 (16SrDNA sequence homologuos to Accession No GU998816) were found to be highly effective in improving the herb and essential oil yield in the cultivars Kushal and Kosi respectively when co-treated with calliterpenone. The results open up the possibilities of using a natural growth promoter along with PGPRs as a bio-agri input for sustainable and organic agriculture.

Tipos de estacas e substratos na propagação vegetativa da menta (Mentha arvensis L. ) / Types of cuttings and substrates in the vegetative propagation of mint (Mentha arvensis L. )

O trabalho teve como objetivo avaliar a propagação vegetativa da menta utilizando diferentes tipos de estacas e substratos. O experimento foi conduzido no Horto de Plantas Medicinais da Unimontes, campus Janaúba - MG. O delineamento experimental utilizado foi o inteiramente casualizado, em esquema fatorial 2 x 4 (dois tipos de estacas e quatro diferentes substratos) com quatro repetições, sendo cada parcela representada por seis estacas. Foram analisadas as variáveis comprimento de parte aérea e de raízes, massa seca de parte aérea e de raízes e número total de brotações formadas por planta. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste de Scott-Knott a 5% de probabilidade. A interação entre os fatores estacas e substratos não foi significativa para as variáveis estudadas, passando-se a estudar o efeito isolado de cada fator. A propagação de Mentha arvensis L. pode ser realizada tanto por estacas apicais como medianas, utilizando o substrato solo + areia + esterco bovino (2:1:1) para a produção de mudas de qualidade.

The purpose of the study was to evaluate the vegetative propagation using different types of mint cuttings and substrates. The experiment was conducted in the Garden of Medicinal Plants of Unimontes, in Janaúba - MG. The experimental design was completely randomized (CRD) in 2 x 4 factorial schemes (two types of poles and four different substrates) with four replications and each plot was represented by six cuttings. The variables analyzed were the length of the shoots and roots, the dry matter of the shoots and roots and the total number of shoots per plant. The data were subject to ANOVA and the means were compared by Scott-Knott's test at 5% of probability. The interaction among stem cuttings and substrates was not significant for the variables studied, thus, the isolated effect of each factor was studied. The propagation of Mentha arvensis L. can be performed either by apical cuttings as medians, using the substrate soil + sand + manure bovine (2:1:1) for the production of quality seedlings.

Nutrient enrichment affects the mechanical resistance of aquatic plants.

For many plant species, nutrient availability induces important anatomical responses, particularly the production of low-density tissues to the detriment of supporting tissues. Due to the contrasting biomechanical properties of plant tissues, these anatomical responses may induce important modifications in the biomechanical properties of plant organs. The aim of this study was to determine the effects of nutrient enrichment on the anatomical traits of two freshwater plant species and its consequences on plant biomechanical performance. Two plant species were grown under controlled conditions in low versus high nutrient levels. The anatomical and biomechanical traits of the plant stems were measured. Both species produced tissues with lower densities under nutrient-rich conditions, accompanied by modifications in the structure of the aerenchyma for one species. As expected, nutrient enrichment also led to important modifications in the biomechanical properties of the stem for both species. In particular, mechanical resistance (breaking force and strength) and stiffness of stems were significantly reduced under nutrient rich conditions. The production of weaker stem tissues as a result of nutrient enrichment may increase the risk of plants to mechanical failure, thus challenging plant maintenance in mechanically stressful or disturbed habitats.

V-cryo-plate procedure as an effective protocol for cryobanks: case study of mint cryopreservation.

A vitrification procedure using aluminium cryo-plates (V-Cryo-plate procedure) was successfully developed and adjusted for in vitro-grown mint (Mentha spp.) shoot tips. Shoots were cultured at 25°C on MS medium containing 0.088 M sucrose for 7 to 14 days after the last subculture. Shoot tips with a basal part (1-1.5 mm × 1 mm) were dissected from the shoots and precultured at 25°C for 1 day on the same medium. Precultured shoot tips were placed on aluminium cryo-plates with 10 wells and embedded in alginate gel. Osmoprotection was performed by immersing the cryo-plates for 30 min at 25 degree C in 25 ml pipetting reservoirs filled with loading solution (2 M glycerol + 0.8 M sucrose). For dehydration, the cryo-plates were transferred and immersed in 25 ml pipetting reservoirs filled with PVS2 for 20 min at 25 degree C. Then the cryo-plates were transferred in uncapped 2 ml cryotubes and directly plunged into liquid nitrogen. For rewarming, shoot tips attached to the cryo-plates were immersed in cryotubes containing 2 ml 1 M sucrose solution at room temperature. Using this procedure, regrowth of cryopreserved shoot tips of line 'Fukuyamajisei' reached over 90 percent. This protocol was successfully applied to 16 additional Mentha lines, with regrowth ranging from 73 percent to 100 percent. This V-Cryo-plate method will facilitate the cryostorage of mint germplasm in our genebank.

Enhanced rosmarinic acid production in cultured plants of two species of Mentha.

In the present investigation an attempt has been made to enhance rosmarinic acid level in plants, grown in vitro, of 2 species of Mentha in presence of 2 precursors in the nutrient media during culture. For in vitro culture establishment and shoot bud multiplication, MS basal media were used supplemented with different concentrations and combinations of different growth regulator like NAA (alpha-napthaleneacetic acid), BAP (6-benzylaminopurine). The medium containing NAA (0.25 mg/L) and BAP (2.5 mg/L) gave the highest potentiality of shoot formation (average 58.0 numbers of shoots) per explant for Mentha piperita L. and the medium containing BAP (2.0 mg/L) gave the highest potentiality of shoot (average 19.2 numbers of shoots) formation per explant for Mentha arvensis L. The complete plants were regenerated in above mentioned media after 8 weeks of subculture. For in vitro enhancement of rosmarinic acid production, the 2 precursors tyrosine (Tyr) and phenylalanine (Phe) were added in the nutrient media at different levels (0.5 mg/L to 15.0 mg/L). Tyrosine was found to be very effective for augmenting rosmarinic acid content in Mentha piperita L. It nearly increased the production up to 1.77 times. In case of Mentha arvensis L., phenylalanine significantly affected the production of rosmarinic acid and the production was nearly 2.03 times more than the control. No significant increase in biomass was observed after addition of these precursors indicating that the added amino acids acting as precursors for rosmarinic acid synthesis were readily utilized in producing rosmarinic acid without promoting growth. Total protein profile also revealed the presence of a specific band in polyacrylamide gel electrophoresis.

Alternativas de propagação na produção de óleo essencial de Mentha canadensis L. no Litoral Norte Catarinense / Propagation alternatives for the production of Mentha canadensis L. essential oil at the Santa Catarina State northern coast

O gênero Mentha é cultivado mundialmente para a produção de óleo essencial, com ênfase no constituinte mentol, amplamente utilizado nas indústrias farmacêutica, cosmética, alimentícia e de higiene pessoal. Este trabalho teve como objetivo avaliar o efeito da utilização de diferentes estruturas de propagação e épocas de colheita de Mentha canadensis L. no Litoral Norte Catarinense. O delineamento experimental foi em blocos ao acaso em esquema fatorial 4x2, comparando quatro estruturas de propagação (estolões com 10, 15 e 20 cm de comprimento e estacas com 5 cm de comprimento) e duas épocas de colheita (95 dias após o plantio e 60 dias após a rebrota). Em cada colheita foi avaliado o crescimento vegetativo e produtividade de óleo essencial e de mentol. Na primeira colheita, houve maior acúmulo de massa seca de folhas, caules e total; produtividade de óleo essencial e de mentol quando o plantio foi realizado com mudas obtidas por estaquia. Na segunda colheita, no entanto, não houve diferença significativa entre os tratamentos para todas as variáveis analisadas. A primeira colheita resultou em médias superiores em relação à segunda colheita à exceção do plantio por estolão com 10 cm em todas as variáveis e estolão com 20 cm para a massa seca de caules. Os constituintes majoritários do óleo essencial foram mentol, mentona e neomentol. Os teores de mentol foram significativamente superiores na segunda colheita e de mentona na primeira colheita, sendo que o neomentol não apresentou diferença significativa em ambas as colheitas. Os resultados obtidos sugerem que a colheita antecipada da rebrota proporciona maior concentração de mentol no óleo essencial. O uso de estolões como estrutura de propagação pode ser considerada uma alternativa viável para menta, pois além de apresentar níveis similares de produção de óleo essencial e de mentol a partir da segunda colheita ao de áreas implantadas com mudas, diminui o custo de produção.

The genus Mentha is cultivated worldwide for essential oil production, with emphasis on its major constituent, menthol, which is used in the pharmaceutical, cosmetics, and food industries and for personal care. This work investigates the use of different propagation structures and harvesting times of Mentha canadensis L., in the Santa Catarina State north coast. The experimental design was in randomized blocks in a 4X2 factorial, comparing four propagation structures (stolon with 10, 15 and 20 cm and stem cuttings with 5 cm) and two harvesting times (95 days after planting and 60 days after the regrowth). In each harvesting time, the vegetative growth, essential oil and menthol productivities were evaluated. The first harvest showed greater leaf, branch and total dry mass accumulation, and essential oil and menthol productivities when stem cuttings were used. In the second harvest, no differences were observed for all evaluated variables. The first harvest resulted in higher averages than the second harvest, with exception in all variables for the use of stolon with 10 cm and in stem dry mass for stolon with 20 cm. The major constituents of the essential oil were menthol, menthone, neomenthol. The levels of menthol were significantly higher in the second harvest and of menthone in the first harvest, while neomenthol showed no significant difference. The results suggest that early harvest of the regrowth results in great menthol concentrations in the essential oil. The use of stolons as a propagation structure can be considered a viable alternative for mint, as it provides similar levels of essential oil and menthol productions after the second harvest, when compared to stem cuttings and also reduces the production cost.

Mentha longifolia in vitro cultures as safe source of flavouring ingredients.

In vitro plantlets and callus of M. longifolia were established and their volatile constituents characterized by GC-MS analysis of their headspaces (HSs) and essential oils (EOs). Significant quali-quantitative differences were found in the aromatic fingerprints in comparison with the M. longifolia parent plants. In fact, limonene and carvone were the main constituents in the EOs of the mother plants, while the aroma of the in vitro plant material were especially enriched in oxygenated terpenes. In particular, huge amounts of piperitenone and piperitenone oxide (75 %) were found for in vitro plantlets, while trans-carvone oxide (19 %) and trans-piperitone epoxide (9 %) were found in callus EO. However, the established in vitro plant material showed lack of pulegone and menthofurane, thus preserving an important feature observed in the volatile fingerprint of the parent plants. In fact, because of their well-known toxicity significant amounts of pulegone and menthofurane may compromise the safety using of mint essential oil. Therefore the in vitro M. longifolia plantlets and callus may be regarded as a potential source of a safe flavouring agent.

Determination of polyphenols in Mentha longifolia and M. piperita field-grown and in vitro plant samples using UPLC-TQ-MS.

Nine polyphenols in the aerial parts of Mentha longifolia have been separated by chromatographic techniques. Their structures have been confirmed by HPLC/electrospray ionization-MS/MS. The compounds identified included rosmarinic acid, salvianolic acid L, dedihydro-salvianolic acid, luteolin-glucuronide, luteolin-diglucuronide, luteolin-glucopyranosyl-rhamnopyranoside, and eriodictyol-glucopyranosyl-rhamnopyranoside. The extracts of M. longifolia and M. piperita field plants, in vitro plants, callus tissues, and cell suspension cultures were profiled, and their polyphenol composition was compared in different tissues and quantified using ultra-performance column liquid chromatography (UPLC)/triple-quadrupole-MS in the selected-ion recording detection mode. Determination of desired compounds was based on calibration curves obtained for standards, which were previously isolated from M. longifolia aerial parts. The UPLC profiles revealed considerable differences in the synthesis of secondary metabolites among samples coming from field plants, in vitro plants, callus tissues, and cell suspension cultures. Plant tissues coming from field cultivation (for both M. piperita and M. longifolia) contained several phenolic compounds (flavonoids and phenolic acids), whereas plants from in vitro conditions, callus tissues, and suspension cultures contained only a few of them. Rosmarinic acid dominated in all of these samples. These results show that under in vitro conditions, the metabolism of phenolics undergoes a fundamental change.

Desenvolvimento vegetativo de Mentha campestris Schur e produção de mentol em diferentes espaçamentos de plantio e épocas de colheita / Vegetative development of Mentha campestris Schur and menthol production in different row spaces and harvest times

A produção de óleos essenciais nas plantas aromáticas é influenciada por fatores bióticos e abióticos. A demanda por esses produtos tem aumentado, sendo os óleos essenciais do gênero Mentha de grande interesse nas indústrias farmacêutica, de cosméticos, alimentícia e agrícola, principalmente em função do composto mentol. Esse trabalho teve como objetivo avaliar o efeito de três espaçamentos de plantio (0,60 x 0,15 m; 0,60 x 0,30 m e 0,60 x 0,45 m) e duas épocas de colheita (60 e 90 dias após o plantio) na espécie Mentha campestris Schur. O experimento foi conduzido no Centro de Estações Experimentais do Canguiri-UFPR, em Pinhais-PR, no período de janeiro a abril de 2008. O delineamento utilizado foi o de blocos ao acaso em esquema de parcelas subdivididas. Houve diferença significativa para todas as variáveis analisadas. As massas secas de folhas, ramos e total foram maiores que na primeira época. Para a biomassa seca de folhas foram observados maiores valores no menor espaçamento de plantio. O rendimento de óleo essencial foi maior na segunda época de colheita e nos espaçamentos maiores. A produtividade do óleo também foi maior na segunda época de colheita, porém no espaçamento mais adensado. Pode-se concluir como recomendação para M. campestris Schur o espaçamento 0,60 x 0,15 m e colheita aos 90 dias, por terem atingido maior biomassa, rendimento de óleo essencial e produtividade de mentol por hectare.

Essential oil production in aromatic plants is influenced by biotic and abiotic factors. The demand for these products has increased, and essential oils from the genus Mentha have been of great interest for pharmaceutical, cosmetic, food and agronomic industries, especially because of the compound menthol. This study aimed to evaluate the effect of three row spaces (0.60 x 0.15 m; 0.60 x 0.30 m and 0.60 x 0.45 m) and two harvest times (60 and 90 days after planting) on the species Mentha campestris Schur. The experiment was carried out at Canguiri Experimental Stations-UFPR, in Pinhais, Paraná State , Brazil, between January and April 2008. The experimental design was in randomized blocks in split-plot. There was significant difference for all variables. In the second harvest time, leaf, branch and total dry yields were higher than those in the first time. For leaf dry mass, the highest values were observed in the narrowest space. Essential oil yield was higher in the second harvest and in wider rows. Essential oil productivity was also higher in the second harvest time, but in the narrowest row. The space 0.60 x 0.15 m and harvest at 90 days can be recommended due to their higher biomass, essential oil yield and menthol productivity per hectare.

Influência de citocininas na micropropagação de Mentha x gracilis Sole / Cytokinin influence on Mentha x gracilis Sole micropropagation

Mentha x gracilis Sole é um híbrido que produz óleos essenciais ricos em monoterpenos. Tendo em vista a propagação clonal desta planta, segmentos nodais provenientes de plantas assépticas, foram cultivados em meio de Murashige e Skoog (MS) suplementado com 0; 0,5; 1,0 e 2,0 µM de cinetina, benzilaminopurina (BAP) ou thidiazuron (TDZ). Após 30 dias, as plantas foram transferidas para meio MS não suplementado com citocinina. Os melhores resultados foram obtidos em meio suplementado com 2 µM de TDZ, mostrando ser método viável para a produção rápida de grande número de mudas. Após a transferência das plantas para a casa de vegetação, as plantas propagadas com TDZ apresentam maior número de tricomas glandulares.

Mentha x gracilis Sole is a hybrid that produces essential oils rich in monoterpenes. Aimed at the clonal propagation of this plant, nodal segments from aseptic plants were cultured in Murashige and Skoog (MSO) medium supplemented with 0; 0.5; 1.0 and 2.0 µM kinetin, benzyl adenine (BAP) or thidiazuron (TDZ). After 30 days, plants were transferred to MOS medium without cytokinin supplementation. The best results were obtained in medium supplemented with 2 µM TDZ, which proved to be a viable method for the rapid production of a large number of seedlings. After transference to the greenhouse, plants propagated with TDZ had a larger number of glandular trichomes.