Compost mixed fruits and vegetable waste biochar with ACC deaminase rhizobacteria can minimize lead stress in mint plants.

High lead (Pb) concentration in soils is becoming a severe threat to human health. It also deteriorates plants, growth, yield and quality of food. Although the use of plant growth-promoting rhizobacteria (PGPR), biochar and compost can be effective environment-friendly amendments for decreasing Pb stress in crop plants, the impacts of their simultaneous co-application has not been well documented. Thus current study was carried, was conducted to investigate the role of rhizobacteria and compost mixed biochar (CB) under Pb stress on selected soil properties and agronomic parameters in mint (Mentha piperita L.) plants. To this end, six treatments were studied: Alcaligenes faecalis, Bacillus amyloliquefaciens, CB, PGPR1 + CB, PGPR2 + CB and control. Results showed that the application A. faecalis + CB significantly decreased soil pH and EC over control. However, OM, nitrogen, phosphorus and potassium concentration were significantly improved in the soil where A. faecalis + CB was applied over control. The A. faecalis + CB treatment significantly improved mint plant root dry weight (58%), leaves dry weight (32%), chlorophyll (37%), and N (46%), P (39%) and K (63%) leave concentration, while also decreasing the leaves Pb uptake by 13.5% when compared to the unamended control. In conclusion, A. faecalis + CB has a greater potential to improve overall soil quality, fertility and mint plant productivity under high Pb soil concentration compared to the sole application of CB and A. faecalis.

Cryopreservation of Plant Shoot Tips of Potato, Mint, Garlic, and Shallot Using Plant Vitrification Solution 3.

Cryopreservation of shoot tips facilitates long-term storage of plant genetic resources which can otherwise only be propagated vegetatively. The vitrification approach using the cryoprotectant plant vitrification solution 3 (PVS3, 50% sucrose and 50% glycerol) is easy to handle, has shown to produce high regrowth percentages in a number of potato, mint, garlic, and shallot accessions, and is, thus, highly suitable for routine cryopreservation of plant genetic resources. In the current chapter, the vitrification procedure is described for potato, mint, garlic, and shallot and includes details about modifications for the different plant species. Special emphasis is given on the preparation of the different culture media, solutions, the culture conditions prior and post-cryopreservation, and the preparation of the shoot tips from different sources. Furthermore, protocols to introduce plants into in vitro culture and methods to estimate cryopreservation success are provided.

Biochar-mediated sequestration of Pb and Cd leads to enhanced productivity in Mentha arvensis.

Immobilization of cadmium (Cd) and lead (Pb) along with the alleviation of their phytotoxicity in Mentha arvensis by biochar was examined in this investigation. A greenhouse experiment was executed to evaluate the effect of biochar (BC) amended Cd and Pb spiked soil on their immobilization and uptake, plant growth, photosynthetic attributes (total chlorophyll, photosynthetic rate, transpiration rate, and stomatal activity) and oxidative enzymes (guaiacol peroxidase: POD; catalase: CAT and superoxide dismutase: SOD). In the present study, the photosynthetic attributes showed that BC significantly improved the total chlorophyll, photosynthetic, transpiration rates, and stomatal activity in the plants. The incorporation of BC in soil increase the Pb and Cd tolerance in M. arvensis vis-à-vis improved the biomass yield and nutrient intake. In addition, biochar has also reduced the POD, CAT, and SOD in the plant as well as improved the soil pH and enzymatic activities. Overall, BC immobilized the Cd and Pb in soil by providing the binding site to the metals and reduced the phytotoxicity in M. arvensis. However, large-scale field trials of BC are required for safe cultivation of M. arvensis which is known for its phytopharmaceuticals importance.

Manganese-induced changes in glandular trichomes density and essential oils production of Mentha aquatica L. at different growth stages.

Production and accumulation of essential oils in plants are influenced by intrinsic and environmental factors. Here, we attempted to elucidate the effect of manganese (Mn) supply on the density of glandular trichomes and the production of essential oils in Mentha aquatica (water mint; syn. Mentha hirsuta Huds.) at the different growth stages. To this aim, plants were treated with 100 µM of Mn (supplied as MnSO4·H2O) at early and late vegetative stages of growth. Then, the control and treated plants were harvested, and biochemical, morphological and molecular analyses indicated that Mn supply has affected M. aquatica at the different growth stages. The biomass, Mn accumulation, glandular trichomes density, essential oils yield and expression levels of the genes encoding enzymes involved in terpenoid biosynthesis pathway (1-Deoxy d-xylulose-5-phosphate synthase (Dxs), geranyl diphosphate synthase (Gpps), isopentenyl diphosphate isomerase (Ippi), ß-caryophyllene synthase (Cps), limonene synthase (Ls) and menthofuran synthase (Mfs)) were increased by Mn supply at both growth stages. However, the increased rates of the assayed parameters were varied between the early and late vegetative stages. Moreover, the content and chemical composition of terpenoid components were affected by Mn supply and plant growth stage. There were positive and weak correlations among the study variables under the Mn supply at the different growth stages. Given these findings, we propose that the application of Mn supply at both early and late vegetative stages elevates the growth, density of glandular trichomes and production of essential oils in M. aquatica.

Manganese modulates the physiological and biochemical responses of Mentha aquatica L. to ultraviolet radiation.

Ultraviolet (UV) radiation as an environmental factor alters the physiological and metabolic processes in plants. Manganese (Mn) is an essential element that is required for plant growth and development. This experiment was conducted in order to determine the effects of Mn supply and UV radiation on the physiological and metabolic responses in Mentha aquatica. With this aim, three levels of Mn and UV treatments were used as follows: basic Hoagland's nutrient solution without UV radiation (control), Mn supply (100µM), UV radiation (2h daily), and UV+100µM Mn. After three weeks of treatments, the root and shoot dry weights and the contents of photosynthetic pigments were decreased under UV radiation condition. However, the contents of flavonoids, soluble carbohydrate, anthocyanins, malonaldehyde (MDA), hydrogen peroxide (H2O2), and the activity of antioxidant enzymes (superoxide dismutase, catalase, and peroxidase) were increased. Interestingly, Mn at 100µM concentration decreased the harmful effects of UV radiation on M. aquatica. In addition, the clear differences were observed in the terpene constituents of M. aquatica after the Mn and UV treatments. In this study, 1, 8-cineole, menthofuran and ß-caryophyllene were the most abundant constituents of essential oils in both the control and treated plants. The correlation analysis between pairs of the primary and secondary metabolites showed that there were positive and negative correlations among the variables under the Mn supply and UV radiation conditions. These findings clearly display a positive effect of external Mn up to 100µM in the nutrient solution on the resistant of M. aquatica to UV radiation.

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.

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.

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.

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.

Chrysolina herbacea modulates terpenoid biosynthesis of Mentha aquatica L.

Interactions between herbivorous insects and plants storing terpenoids are poorly understood. This study describes the ability of Chrysolina herbacea to use volatiles emitted by undamaged Mentha aquatica plants as attractants and the plant's response to herbivory, which involves the production of deterrent molecules. Emitted plant volatiles were analyzed by GC-MS. The insect's response to plant volatiles was tested by Y-tube olfactometer bioassays. Total RNA was extracted from control plants, mechanically damaged leaves, and leaves damaged by herbivores. The terpenoid quantitative gene expressions (qPCR) were then assayed. Upon herbivory, M. aquatica synthesizes and emits (+)-menthofuran, which acts as a deterrent to C. herbacea. Herbivory was found to up-regulate the expression of genes involved in terpenoid biosynthesis. The increased emission of (+)-menthofuran was correlated with the upregulation of (+)-menthofuran synthase.

Synthesis of a novel plant growth promoter from gallic acid.

Gallic acid has been modified to naphthophenone derivatives with esterified fatty acid side chain. Compound 12, an ethyl crotonate ester of naphthophenone derivative has shown potent auxin like growth promoter activity. This is the first example of naphthophenone derivatives with plant growth promoting activity.

Influence of pH on lead uptake, chlorophyll and nitrogen content of Nasturtium officinale R. Br. and Mentha aquatica L.

In the present study, effects of pH (5.0, 7.0 and 9.0) and lead (1, 5, 10, 25, 50 and 100 microg mL(-1)) were investigated on uptake of lead, content of chlorophyll and nitrogen in Nasturtium officinale and Mentha aquatica. Total chlorophyll and nitrogen contents were adversely affected from Pb2+ concentrations dose dependently at each pH. The macrophytes were adversely affected by pH 5.0 or more than 9.0. After 12-days Pb2+ treatment, results showed that lead accumulation of macrophyte tissues was variable. According to the parts of the macrophytes, Pb2+ amounts were generally found at all tested pH levels and the metal concentrations for M. aquatica in the following order: root > stem > leaf and for N. officinale root > leaf > stem. Pb2+ concentrations in plant tissues (root, stem, leaf) in relation to pH were generally found for both macrophytes in following order: 7.0 > 9.0 > 5.0. Despite of the fact that high Pb2+ accumulation was observed in root tissues of the macrophytes, low metal accumulation was measured in the above-ground parts indicating low root-leaf translocation. The study indicated that uptake rate of Pb2+ and its toxicity on Chlorophyll and nitrogen contents in the macrophytes were dependent upon pH value of solutions.

Effect of clinorotation on in vitro cultured explants of Mentha piperita L.

An in vitro culture system was used to study the influence of gravity on axillary shoot formation and adventitious root regeneration in Mentha piperita L. The direction of the gravity vector was altered by displacing stem node explants in different orientations. Also, microgravity conditions were simulated by rotating the explants on a horizontal clinostat so that the main axis of nodes was either parallel (Cpa) or perpendicular to the clinostat axis (Ccp and Ccf, centripetally and centrifugally oriented, respectively). Mint nodes were cultured on solidified Linsmaier and Skoog's medium [Physiol. Plant. 18 (1965) 100] adding a filter-sterilized aqueous solution of 2 mg/l benzyladenine (BA) in half of the cultures. The proliferation of axillary shoots as well as adventitious root formation were not affected by altering upright explant orientation. On the contrary clinorotation was able to modify plantlet development. In absence of BA, leaf width was hindered by Cpa treatment and penultimate internode length was enhanced by Ccp. Furthermore, a negative effect of Cpa treatment was observed in root length parameter, while Ccp increased the root number both in absence and in presence of BA. An effect strictly connected to clinorotation in presence of BA was the occurrence of hyperhydricity. Moreover, explants under clinorotation treatments switched their gravitropic response modifying shoot curvature.