Cyanobacterial elicitor enhances the biomass of Mentha piperita L. and improves the production of high-value rosmarinic acid under in vitro culture of apical meristem.

BACKGROUND: Rosmarinic acid (RA), like other phenolic compounds, is sources of antioxidants and anti-inflammatory agents in medicinal plants. In vitro culture of plants can improve the medicinal plants' metabolite profile and phenolic compound quantity. To date, various methods have been proposed to increase this medicinal metabolite in plants, among which the use of bioelicitors can be mentioned. In the present study, a native isolate of heterocystous cyanobacteria, Nostoc spongiaeforme var. tenue ISB65, was used to stimulate the production of biomass and content of RA in Mentha piperita L. (peppermint) grown in vitro from apical meristem. Mentha piperita L. explants were inoculated in half strength Murashige and Skoog (1/2 MS) medium containing cyanobacterial lysate (CL). After 50 days of culturing, the growth indices, the content of photosynthetic pigments, and RA in control and treated plants were measured. RESULTS: CL inoculation resulted in a significant enhancement in the vegetative growth indices of peppermint, including root and shoot length, plant biomass and leaf number. The content of photosynthetic pigments also increased in cyanobacteria-treated plants. Inoculation with CL increased the RA content by 2.3-fold, meaning that the plants treated with CL had the highest RA content (7.68 mg. g- 1 dry weight) compared to the control (3.42 mg. g- 1 dry weight). Additionally, HPLC analysis revealed the presence of several auxins in CL. CONCLUSIONS: The presence of auxins and the chemical content of CL such as K+ and Ca2+, as regulators of metabolic pathways and molecular activities of cells, may be responsible for the enhanced growth and phenolic compounds of plants under tissue culture conditions. An improvement in RA content in the tissue culture of medicinal plants treated with CL was reported for the first time in this investigation.

Does co-inoculation of mycorrhiza and Piriformospora indica fungi enhance the efficiency of chlorophyll fluorescence and essential oil composition in peppermint under irrigation with saline water from the Caspian Sea?

Symbiotic associations with endophytic fungi are ecologically important for medicinal and aromatic plants. Endophytic fungi highly affect the quantity and quality of herbal products. In this study, a pot experiment was carried out in the greenhouse to investigate the interactive effects of Piriformospora indica and arbuscular mycorrhizal (AMF) inoculation on the chlorophyll fluorescence, essential oil composition, and antioxidant enzymes of peppermint under saline condition. The results showed that Fo, YNPQ, YNO, and NPQ values were obviously increased under salinity conditions, while essential oil content, chlorophyll a and b, gs, Fm, Fv, ETR, ФPSII and Fv/Fm ratio decreased by increasing salinity. In addition, salt induced the excess Na+ uptake, whereas the opposite trend was observed for P and K+. The synergistic association of P. indica and AMF caused a considerable increase in the antioxidant ability, essential oil content, Fv/Fm ratio, ФPSII, and amount of P and K+ uptake in salt-stressed plants. The main peppermint oil constituents, menthol, menthone, and 1,8-cineole increased considerably in inoculated plants. Besides, the applied endophytic fungi positively enhanced the ability of peppermint to alleviate the negative effect of the salinity stress.

Improving Phenolic Total Content and Monoterpene in Mentha x piperita by Using Salicylic Acid or Methyl Jasmonate Combined with Rhizobacteria Inoculation.

The effects of plant inoculation with plant growth-promoting rhizobacteria (PGPR) and those resulting from the exogenous application of salicylic acid (SA) or methyl jasmonte (MeJA) on total phenolic content (TPC) and monoterpenes in Mentha x piperita plants were investigated. Although the PGPR inoculation response has been studied for many plant species, the combination of PGPR and exogenous phytohormones has not been investigated in aromatic plant species. The exogenous application of SA produced an increase in TPC that, in general, was of a similar level when applied alone as when combined with PGPR. This increase in TPC was correlated with an increase in the activity of the enzyme phenylalanine ammonia lyase (PAL). Also, the application of MeJA at different concentrations in combination with inoculation with PGPR produced an increase in TPC, which was more relevant at 4 mM, with a synergism effect being observed. With respect to the main monoterpene concentrations present in peppermint essential oil (EO), it was observed that SA or MeJA application produced a significant increase similar to that of the combination with rhizobacteria. However, when plants were exposed to 2 mM MeJA and inoculated, an important increase was produced in the concentration on menthol, pulegone, linalool, limonene, and menthone concentrations. Rhizobacteria inoculation, the treatment with SA and MeJA, and the combination of both were found to affect the amount of the main monoterpenes present in the EO of M. piperita. For this reason, the expressions of genes related to the biosynthesis of monoterpene were evaluated, with this expression being positively affected by MeJA application and PGPR inoculation, but was not modified by SA application. Our results demonstrate that MeJA or SA application combined with inoculation with PGPR constitutes an advantageous management practice for improving the production of secondary metabolites from M. piperita.

Survival of Salmonella on chamomile, peppermint, and green tea during storage and subsequent survival or growth following tea brewing.

The survival of Salmonella on dried chamomile flowers, peppermint leaves, and green tea leaves stored under different conditions was examined. Survival and growth of Salmonella was also assessed after subsequent brewing using dried inoculated teas. A Salmonella enterica serovar cocktail was inoculated onto different dried tea leaves or flowers to give starting populations of approximately 10 log CFU/g. The inoculum was allowed to dry (at ambient temperature for 24 h) onto the dried leaves or flowers prior to storage under 25 and 35 °C at low (<30% relative humidity [RH]) and high (>90% RH) humidity levels. Under the four storage conditions tested, survival followed the order 25 °C with low RH > 35 °C with low RH > 25 °C with high RH > 35 °C with high RH. Salmonella losses at 25 °C with low RH occurred primarily during drying, after which populations showed little decline over 6 months. In contrast, Salmonella decreased below detection after 45 days at 35 °C and high RH in all teas tested. The thermal resistance of Salmonella was assessed at 55 °C immediately after inoculation of tea leaves or flowers, after drying (24 h) onto tea leaves or flowers, and after 28 days of storage at 25 °C with low RH. All conditions resulted in similar D-values (2.78 ± 0.12, 3.04 ± 0.07, and 2.78 ± 0.56, at 0 h, 24 h, and 28 days, respectively), indicating thermal resistance of Salmonella in brewed tea did not change after desiccation and 28 days of storage. In addition, all brewed teas tested supported the growth of Salmonella. If Salmonella survives after storage, it may also survive and grow after a home brewing process.

[Detection of fungal metabolites showing toxic activity through Artemia salina bioassay]. / Detección de metabolitos fúngicos con actividad tóxica mediante bioensayo sobre Artemia salina.

The aim of this study was to detect toxic metabolites from fungi contaminating food and medicinal herbs by applying the toxicity assay to Artemia salina. According to toxicity percentages, the extracts were classified as nontoxic (NT), slightly toxic (ST), toxic (T) and highly toxic (HT). Those classified as T and HT were assayed for mycotoxins. Only 6 out of 71 strains were found to be T (8.5%) for A. salina. Penicillium brevicompactum Dierckx, isolated from sausages, was found to be HT, mainly due to the presence of ochratoxin A and two other unidentified metabolites.

A comparison of selected media and incubation temperatures for isolation of microscopic fungi from dried medicinal plants.

The study compared selected media and incubation temperatures for isolation of fungi from dried medicinal plants (chamomile, peppermint, lemon balm, St. John's wort and two herbal mixtures). The DG18 medium was found to be the most suitable for characterization of the mycoflora at 25 degrees C. The medium selection for 37 degrees C was dependent on the species to be isolated. MEA + 40% sacharose and YpSs were found to be the best media for isolation of thermophilic and thermotolerant fungi from dried medicinal plants.

Occurence of fungal diseases on lemon balm (Mellisa officinalis L.) and peppermint (Mentha x piperita L.) in the region of Malopolska.

This paper presents the results of the researches carried out on the subject of the diseases on herbs such as lemon balm and peppermint focusing on the health status of the plants grown in the region of Malopolska. The field and laboratory research proved that perpetrators of the diseases on the examined plants were fungi species with the numerical majority. On lemon balm septoria leaf spot (Septoria melissae) was most often observed. Moreover, fungal genera of different taxonomic groups were detected. Alternaria, Epicoccum, Fusarium, and Sclerotinia sclerotiorum occurred most often on the medicinal plant samples. The disease mint rust Puccinia menthae, has caused major problems for the peppermint growers. In the populations of fungi found on the diseased leaves the dominating were Alternaria, Epicoccum and Sphaceloma menthae. It has been stated that among all fungi isolated from lemon balm, the species Fusarium avenaceum had the highest pathogenicity to seedlings (80% of diseased seedlings). For peppermint plants the highest pathogenicity had species Epicoccum purpurascens, and caused 82% of diseased seedlings. Conducted evaluation of health status of plants showed that the lemon balm mean disease index was yearly differentiated and was the highest in 2003 (44.39). For peppermint, the highest disease index was in third year of cultivation (62.75) and was statistically higher than in previous years.

Effect of an arbuscular mycorrhizal fungus, Glomus mosseae, and a rock-phosphate-solubilizing fungus, Penicillium thomii, on Mentha piperita growth in a soilless medium.

Rock phosphate effect on English mint (Mentha piperita L.) grown on steamed perlite:vermiculite (1:1, v:v) substrate, with and without rock phosphate, was evaluated in greenhouse experiments. Five treatments were carried out by inoculation with an arbuscular mycorrhizal fungus Glomus mosseae and a phosphorus solubilizing microorganism Penicillium thomii. Plant aerial biomass, phosphorus concentration in plant tissue, and P available in the substrate, were evaluated upon two harvests. After the first harvest, plant aerial biomass did not show significant differences between treatments using rock phosphate as fertilizer, although P content in plants inoculated with P. thomii was higher. The second harvest revealed a higher biomass and plant tissue P content in treatments inoculated with G. mosseae. P. thomii increased P available in the substrate, whereas in the absence of G. mosseae, it did not enhance plant tissue P content. Mycorrhizal colonization was not affected by P. thomii. Microbial inoculation effect on English mint growth was also evaluated. The microbial effect was positive in all treatments when compared with the control without rock phosphate.

[Study on the quantitative and qualitative composition of fungi in dried medicinal plants]. / Badania skladu ilosciowego i jakosciowego grzybów w suszach roslin leczniczych.

The quantitative and qualitative composition of fungi was determined in selected dried medicinal plants purchased in one of the herbal shops in Szczecin, Poland. The samples examined were as follows: chamomile (Flos Chamomillae), peppermint (Folium Menthae piperitae), lemon balm (Folium Melissae), St. John's wort (Herba Hyperici), and two herbal mixtures. The fungal composition depended on the specified sample. Xerophilic fungi, i.e. Eurotium amstelodami, E. herbariorum, E. rubrum and Wallemia sebi were isolated from dried medicinal plants. E. amstelodami was the predominating species. The prevailing thermophilic and thermotolerant species were Rhizopus microsporus var. rhizopodiformis and Aspergillus fumigatus. Pink and white yeasts were also numerous in some samples. Except for Aspergillus niger, mesophilic and toxigenous species were found to occur infrequently in the samples. However, Aspergillus versicolor was found to occur abundantly in lemon balm.

Exposure to airborne microorganisms, dust and endotoxin during processing of peppermint and chamomile herbs on farms.

The aim of this study was to determine the levels of microorganisms, dust and endotoxin in the air during processing of peppermint (Mentha piperita) and chamomile (Matricaria recutita) by herb farmers, and to examine the species composition of airborne microflora. Air samples were collected on glass fibre filters by use of personal samplers on 13 farms owned by herb cultivating farmers, located in Lublin province (eastern Poland). The concentrations of total viable microorganisms (bacteria + fungi) in the farm air during processing of peppermint herb were large, within a range from 895.1-6,015.8 x 10(3) cfu/m(3) (median 1,055.3 x 10(3) cfu/m(3)). During processing of chamomile herb they were much lower and varied within a range from 0.88-295.6 x 10(3) cfu/m(3) (median 27.3 x 10(3) cfu/m(3)). Gram-negative bacteria distinctly prevailed during processing of peppermint leaves, forming 46.4-88.5 % of the total airborne microflora. During processing of chamomile herb, Gram-negative bacteria were dominant at 3 out of 6 sampling sites forming 54.7-75.3 % of total microflora, whereas at the remaining 3 sites the most common were fungi forming 46.2-99.9 % of the total count. The species Pantoea agglomerans (synonyms: Erwinia herbicola, Enterobacter agglomerans ), having strong allergenic and endotoxic properties, distinctly prevailed among Gram-negative isolates. Among fungi, the most common species was Alternaria alternata. The concentrations of airborne dust and endotoxin determined on the examined herb farms were large. The concentrations of airborne dust during peppermint and chamomile processing ranged from 86.7-958.9 mg/m(3), and from 1.1-499.2 mg/m(3), respectively (medians 552.3 mg/m(3) and 12.3 mg/m(3)). The concentrations of airborne endotoxin determined during peppermint and chamomile processing were within a wide range 1.53-208.33 microg/m(3) and 0.005-2604.19 microg/m(3) respectively (medians 57.3 microg/m(3) and 0.96 microg/m(3)). In conclusion, farmers cultivating peppermint are exposed during processing of this herb to large concentrations of airborne microorganisms, dust and endotoxin posing a risk of work-related respiratory disease. The exposure to bioaerosols during processing of chamomile is lower; nevertheless, peak values create a respiratory risk for exposed farmers.