Hybridization in the section Mentha (Lamiaceae) inferred from AFLP markers

The amplified fragment length polymorphism (AFLP) method was used to evaluate genetic diversity and to assess genetic relationships within the section Mentha in order to clarify the taxonomy of several interspecific mint hybrids with molecular markers. To this end, genetic diversity of 62 Mentha accessions from different geographic origins, representing five species and three hybrids, was assessed. Three EcoRI/MseI AFLP primer combinations generated an average of 40 AFLP markers per primer combination, ranging in size from 50 to 500 base pairs (bp). The percentage of markers polymorphic ranged from 50% to 60% across all accessions studied. According to phenetic and cladistic analysis, the 62 mint accessions were grouped into two major clusters. Principal coordinates analysis separated species into well-defined groups, and clear relationships between species and hybrids could be described. Our AFLP analysis supports taxonomic classification established among Mentha species by conventional (morphological, cytological, and chemical) methods. It allows the assessment of phenetic relationships between species and the hybrids M. spicata and M. × piperita, largely cultivated all over the world for their menthol source, and provides new insights into the subdivision of M. spicata, based for the first time on molecular markers.     

Enhanced nutrient uptake in salt-stressed Mentha piperita using magnetically treated water

The improvement of the growth and quality of medicinal plants under stress is of significance, worldwide. The hypothesis was to alleviate salinity stress in Mentha piperita by enhancing nutrient uptake using magnetically treated water, which to our knowledge has not been previously investigated. The objective was to test the effects of magnetized water (using alternating magnetic fields) (main plots, M1-M4 representing control, 100, 200, and 300 mT, respectively), salinity (subplots, S1-S4 representing control, 40, 80, and 120 mM NaCl, respectively), and growth medium (sub-subplots, X1-X4 representing coco peat, palm, coco peat + perlite, and palm + perlite, respectively) on M. piperita nutrient uptake in the greenhouse. The M treatments, especially the 100 and 200 mT levels, significantly increased plant N (1.08%, S3M4X1), P (0.89%, S3M3X1), K (3.23%, S3M3X1), Ca (53.6 mg/kg, S4M4X4), and Mg (39.63 mg/kg, S3M3X2) concentrations (compared with control at 0.71, 0.49, 2.4, 26.63, 1.63) even at the highest level of salinity. Magnetically treated water also significantly enhanced plant Fe and Zn concentration to a maximum of 750 μg/kg (M4S3X1) and 94.67 μg/kg (S4M4X3), under salinity stress, respectively. The single and the combined use of organic and mineral media significantly affected plant nutrient uptake, especially when used with the proper rate of M treatment. If combined with the proper growth medium, the magnetized water may be more effective on the alleviation of salt stress in Mentha piperita by enhancing nutrient uptake.

     

Medicinal and aromatic plant materials as nitrification inhibitors for augmenting yield and nitrogen uptake of Japanese mint (Mentha arvensis L. Var. Piperascens)

Pot experiments were conducted to evaluate the relative performance of medicinal and aromatic plant materials and dicyandiamide (DCD) as nitrification inhibitors to regulate transformation of N from urea. Their effect on the efficiencies of use of N by Japanese mint (Mentha arvensis cv. Hy 77) was tested. Urea was coated with these materials viz., Mentha spicata, Artemisia annua or DCD at the rate of 5% (w/w) of fertilizer urea using an appropriate coating technique. Nimin (tetranortriterpenoids, an ethanol extract of neem (Azadirachta indica Juss) coating was done at the rate of 1% w/w of urea. Fertilizer nitrogen was applied at 100 and 200 mg kg(-1) soil. These natural coating materials significantly increased the herb and essential oil yields of the crop at both rates of fertilizer nitrogen compared to urea alone and were found to be as effective as DCD in retarding NO3- formation in soil. Herb yield increased by 6-81% when compared to uncoated urea. The increase in essential oil yield ranged between 3% and 68% due to coating. The effectiveness of the nitrification-inhibitor--coated urea, however, varied with the soils used and the rate of fertilizer nitrogen applied. The results suggest that the natural products could be potential nitrification inhibitors for increasing fertilizer N use efficiency.     

Inhibition by the essential oils of peppermint and spearmint of the growth of pathogenic bacteria

The effects of the, essential oils of peppermint (Mentha piperita L.), spearmint Mentha spicata L.) and Japanese mint (Mentha, arvensis L.), of four major constituents of the esssential oil of peppermint, and of three major constituents of the essential oil of spearmint, on the proliferation of Helicobacter pylori, Salmonella enteritidis, Escherichia coli O157:H7, methicillin-resistant Staphylococcus aureus (MRSA) and methicillin sensitive Staphylococccus aureus (MSSA) were examined. The essential oils and the various constituents inhibited the proliferation of each strain in liquid culture in a dose-dependent manner. In addition, they exhibited bactericidal activity in phosphate-buffered saline. The antibacterial activities varied among the bacterial species tested but were almost the same against antibiotic-resistant and antibiotic-sensitive strains of Helicobacter pylori and S. aureus. Thus, the essential oils and their constituents may be useful as potential antibacterial agents for inhibition of the growth of pathogens.     

The origin of mentha — gracilis (Lamiaceae). II. essential oils

Essential oils were eamined in nine clones of Mentha arvensis, four clones of M. spicata, and 20 clones of M. gracilis. An F₁ hybrid of M. arvensis M. spicata, selected on the basis of morphology and chromosome number, was matched with one clone of M. Gracilis. Genes for the inheritance of limonene, 1,8-cineole, linalool, isomenthone, carvone, and piperitenone oide were identified in one clone of M. arvensis and two clones of M. spicata. The range of essential oil compounds detected indicates that no one character can be used to identify M. gracilis, but the critical compounds of the oil of M. gracilis can be derived from crosses of M. arvensis M. spicata.     

Use of RAPD and AFLP markers to identify inter- and intraspecific hybrids of Mentha

Three controlled crosses were carried out involving Mentha arvensis and Mentha spicata [M. spicata CIMAP/C30 x M. spicata CIMAP/C33 (cv. Neera); M. arvensis CIMAP/C18 x CIMAP/C17 (cv. Kalka); and M. arvensis CIMAP/C17 x M. spicata CIMAP/C33]. The parents were subjected to random amplified polymorphic DNA (RAPD) analysis with 80 primers, and polymorphic primers were tested for detecting coinherited RAPD profiles among the progeny of these crosses. Of 50 seedlings tested from each intraspecific cross, all demonstrated dominant profiles with the selected RAPD primers except the detected hybrid from respective crosses. Coinherited markers could be detected with the primers OPJ 01, MAP 06, OPT 08, and OPO 20 for M. arvensis; OPJ 05, OPJ 14, OPO 19, and OPT 09 for M. spicata; and OPJ 07, OPJ 10, OPJ 11, OPJ 14, and OPO 02 for the cross M. arvensis x M. spicata. In our amplified fragment length polymorphism (AFLP) analysis, 40 coinherited marker fragments were identified for the cross involving M. arvensis, 32 for the cross involving M. spicata, and 41 for the interspecific cross between M. arvensis and M. spicata. In all crosses, similarity values between the parents were less than those between the parents and the hybrids. Although RAPD markers are generally considered dominant, it is possible to identify a few codominant markers that behave like restriction fragment length polymorphism (RFLP) markers. This molecular marker system may be helpful in rapidly screening out hybrids in crops where cross-pollination is a problem.     

Effect of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus fasciculatum on the essential oil yield related characters and nutrient acquisition in the crops of different cultivars of menthol mint (Mentha arvensis) under field conditions.

The effects of inoculation with vesicular-arbuscular mycorrhizal (VAM) fungus Glomusfasciculatum on the root colonization, growth, essential oil yield and nutrient acquisition of three cultivars of menthol mint (Mentha arvensis); Kalka, Shivalik and Gomti, were studied under field conditions. The VAM inoculation significantly increased the root colonization, plant height, fresh herbage and dry matter yield. oil content and oil yield as compared to non-inoculated cultivars. The effect of VAM inoculation on the root colonization, growth and yield of mint was more pronounced with the cv Shivalik than the cvs Kalka and Gomati, indicating Shivalik as a highly mycorrhizal dependent genotype. VAM inoculation significantly increased the uptake of N, P and K by shoot tissues of mint, but most markedly increased the uptake of P. The VAM-inoculated mint plants depleted the available N, P and K in the rhizosphere soil as compared to non-inoculated control plants, however the extent of nutrient depletion was greater for P than N and K. We conclude that the VAM inoculation could significantly increase the root colonization, growth, essential oil yield and nutrient acquisition of mint for obtaining economic production under field conditions.     

鸡内金中氨基酸及营养元素含量的测定

采用日立 835 - 5 0型氨基酸自动分析仪和岛津Z - 80 0 0型原子吸收分光光计。测定了鸡内金中氨基酸和营养元素的含量 ,结果表明 :鸡内金中营养元素钾、钙、镁、铜、锌、铁、锰含量分别为 36 9.5 0、12 0 .5 6、470 .77、31.89、32 .42、5 5 9.6 4、8.11mg/kg ;氨基酸总量为 86 .92 4% ,必需氨基酸占氨基酸总量的 30 .2 6 4%。     

Effects of acute oral lead exposure on the levels of essential elements of mice: a metallomics and dose-dependent study

Background and objectivesLead (Pb) has been reported to disturb the metabolism of essential elements, such as calcium (Ca), magnesium (Mg), iron (Fe) and zinc (Zn) in vivo. This study focused on the relationship between various dose of Pb and the essential elements.Methods50 healthy male C57BL/6 mice underwent oral administration of 0.2 mL lead acetate trihydrate solution (0, 20, 100, 500, and 1000 mg Pb/day/kg body weight) for 3 days. The concentrations of Pb and four essential elements (Ca, Zn, Fe and Mg) in the blood, kidney, liver, bone and brain were quantified with inductively coupled plasma mass spectrometry.ResultsVarious doses of Pb led to significant increases in the contents of Ca, Fe and Zn in the liver, and decreased contents of Mg and Fe in the blood in a dose-dependent pattern. The Pb dose of 20 mg/kg reduced the concentration of bone Ca, which did not continue to show an obvious decline with continued increases in the oral Pb dose. Pb also caused alterations in the Mg distribution pattern, and decreased the correlation of Mg, Ca and Zn in the brain, both findings were dose-dependent. In addition to the changes in metallomics, the related oxidative stress was exacerbated, but no significant changes were detected in hepatic and renal histopathological lesions after a short period of Pb exposure.ConclusionsThis study contributes to a thorough analysis of the Pb-poisoning mechanism, and indicates that the concentrations of essential elements could be used as sensitive toxicological indicators of Pb exposure.     

Organization of monoterpene biosynthesis in Mentha. Immunocytochemical localizations of geranyl diphosphate synthase, limonene-6-hydroxylase, isopiperitenol dehydrogenase, and pulegone reductase

We present immunocytochemical localizations of four enzymes involved in p-menthane monoterpene biosynthesis in mint: the large and small subunits of peppermint (Mentha x piperita) geranyl diphosphate synthase, spearmint (Mentha spicata) (-)-(4S)-limonene-6-hydroxylase, peppermint (-)-trans-isopiperitenol dehydrogenase, and peppermint (+)-pulegone reductase. All were localized to the secretory cells of peltate glandular trichomes with abundant labeling corresponding to the secretory phase of gland development. Immunogold labeling of geranyl diphosphate synthase occurred within secretory cell leucoplasts, (-)-4S-limonene-6-hydroxylase labeling was associated with gland cell endoplasmic reticulum, (-)-trans-isopiperitenol dehydrogenase labeling was restricted to secretory cell mitochondria, while (+)-pulegone reductase labeling occurred only in secretory cell cytoplasm. We discuss this pathway compartmentalization in relation to possible mechanisms for the intracellular movement of monoterpene metabolites, and for monoterpene secretion into the extracellular essential oil storage cavity.     

A unique interspecific hybrid spearmint clone with growth properties of Menthaarvensis L. and oil qualities of Mentha spicata L.

Crossing blocks of Mentha arvensis cv Kalka (menthol mint without carvone) and Mentha spicata cv Neera (carvone mint without menthol) in alternate rows (2:1::Kalka:Neera), and pollination of florets of Kalka with that from Neera, yielded a carvone-rich variant among the open-pollinated seedlings. The variant possesses the hybrid phenotype, including the vigorous M. arvensis growth habit and the synthesis of rich oil aroma supplemented with a menthol tinge (carvone 64%–76% against 58% for the normal carvone mint cv Neera), and thus a novel combination of the essential oil. Chromosome counts and random amplified polymorphic DNA analysis confirmed that this spearmint variant, designated as the variety Neerkalka, is a unique interspecific hybrid (2n=5x=60) of M. arvensis cv Kalka (2n=8x=96) and M. spicata cv Neera (2n=2x=24). Vegetative multiplication of the hybrid was facilitated by its underground sucker-reproducing ability which is otherwise absent in spearmints. The per cent improvement in the variant ranged from 31–97 for herbage yield and 95–317 for oil yield over the standard spearmint varieties (MSS-5, Arka and Neera), with per hectare oil yields of 125.0 kg, 139.0 kg and 65.0 kg, respectively. Received: 25 March 2000 / Accepted: 3 July 2000     

Mineral concentrations in whole mice and rats used as food

Calcium (Ca), magnesium (Mg), copper (Cu), iron (Fe), manganese (MD), and zinc (Zn) concentrations were measured in whole mice (five sizes) and rats (six sizes). Ca concentrations increased with age in both mice (1.2–2.3%) and rats (1.9–3.3%). Mg levels ranged from 0.09 to 0.13% (mice) and from 0.11 to 0.18% (rats), with the medium-size class of both species having the highest values. Cu (7.9–19.2 and 10.8–60.6 mg/kg) and Zn (58.0–82.5 and 73.2–113.6 mg/kg) generally decreased with age in both species, while Mn levels tended to increase with age (0.2–13.1 mg/kg, mice; 6.2–28.3 mg/kg, rats). Fe values were highest in neonates and adult size classes, ranging from 113.4 to 181.3 mg/kg (mice) and 111.3 to 332.6 mg/kg (rats). Rats usually contained higher concentrations of individual minerals than equivalent age categories of mice, even though both were fed identical diets. All rodents analyzed generally met known dietary requirements of mammalian carnivores, but differences between mice and rats were apparent. Specific mineral nutrient requirements for carnivorous birds and reptiles have not been determined. © 1996 Wiley-Liss, Inc.     

Studies of Zn, Mg, Cu, Ca and Fe in cadmium poisoned toads before and after treatment with EDTA

Zn, Mg, Cu, Ca and Fe were determined spectrophotometrically in liver, kidneys, muscle, spleen and blood of Bufo regularis after a single i.m. injection of 6.2 mg Cd/kg (which represents the 96 hr LD50) alone or in combination with 40 mg EDTA/kg (the minimal EDTA concentration causing 100% survival over that period). Cadmium administration caused recognizable effects on the essential metals levels in different tissues and organs. In the majority of the tissues and organs studied, zinc and copper concentrations returned to their normal ranges in animals that received both cadmium and EDTA. In contrast, magnesium, calcium and iron contents not only returned back to their control values but also exceeded them.     

Protective effects of solvent fractions of Mentha spicata (L.) leaves evaluated on 4-nitroquinoline-1-oxide induced chromosome damage and apoptosis in mouse bone marrow cells

Spearmint leaves (Mentha spicata L.) contain high levels of antioxidants that are known to protect against both exogenous and endogenous DNA damage. In this study, the protective effects of the hexane fraction (HF), chloroform fraction (CF) and ethyl acetate fraction (EAF) in an ethanol extract from M. spicata were evaluated against 4-nitroquinoline-1-oxide (4-NQO) induced chromosome damage and apoptosis in bone marrow cells of Swiss albino mice. Two (EAF; 80 and 160 mg/ kg body weight - bw) or three (HF and CF; 80, 160 and 320 mg/ kg bw) doses of solvent fractions or vehicle control (25% DMSO in water) were administered orally for five consecutive days. Upon the sixth day, 4-NQO was injected intraperitoneally. The animals were killed the following day. Other control groups were comprised of animals treated with either the vehicle control or the various doses of solvent fractions, but with no 4-NQO treatment. 4-NQO induced micro-nucleated polychromatic erythrocytes (MnPCEs) in all the test groups. However, pre-treatment of animals with the solvent fractions significantly reduced the 4-NQO-induced MnPCEs as well as the percentage of apoptotic cells. The reduction of both MnPCE and apoptosis was more evident following the pre-treatment of animals with 160 mg/kg bw EAF.     

Essential Oils as Potent Source of Nematicidal Compounds

Bioassay tests were conducted to find out the nematicidal activity of eight essential oils against Meloidogyne incognita (Kofoid and White) Chitwood at four concentrations. Maximum activity was recorded in oils of Eucalyptus citriodora , Eucalyptus hybrida and Ocimum basilicum followed by Pelargonium graveolens , Cymbopogon martinii, Mentha arvensis, Mentha piperita and Mentha spicata oils, respectively. The eucalyptus ( E. citriodora and E. hybrida ) and Indian basil ( O. basilicum ) oils were highly toxic to M. incognita even at the lower concentrations, namely 500 and 250 ppm. The remaining oils were also toxic to the nematode but at different amounts.     

High Urinary Iodine Concentration Among Breastfed Infants and the Factors Associated with Iodine Content in Breast Milk

Cadmium (Cd) is a highly toxic element, which may cause toxicity to most organs in the body. Zinc (Zn) and magnesium (Mg) are essential minerals with probable benefits on Cd harmful effects. Finding an efficient and non-pathological treatment against Cd toxicity seems promising. Fifty adult rats were divided into ten experimental groups of five rats each. The Cd group was treated with 1 mg Cd/kg and the control group received 0.5 cm³ normal saline. The other eight groups received Zn (0.5 and 1.5 mg/kg) and Mg (0.5 and 1.5 mg/kg) either alone or in combination with 1 mg Cd/kg through IP injection for 3 weeks. Testis malondialdehyde (MDA), sperm parameters, and testis histopathology were investigated. Cd reduced sperm parameters and increased testis MDA. Moreover, Cd exposure caused a significant histological damage in testis of male rats. However, Zn or Mg treatment prevented and reversed Cd toxic alterations in testis. These findings suggest that co-administration of Zn or Mg could improve cadmium testicular toxicity in male Wistar rats.     

Mentha spicata (Lamiaceae) chemotypes growing wild in Greece

Mentha spicata is the commonest mint species growing wild in Greece, exhibiting great morphological and chemical variability. The oil content from different wild populations examined ranged from 0.3% to 2.2%; the most common value being ca.1%. Though commercially exploited M. spicata plants are always rich in carvone and dihydrocarvone, wild populations are very variable; four different chemotypes were distinguished within the species. These chemotypes are characterized by the high contribution of the following compounds: (1) linalool, (2) piperitone oxide or piperitenone oxide, (3) carvone-dihydrocarvone and (4) pulegone-menthone-isomenthone. This chemical variability suggests the possibility of further developing and exploiting the full potential of the species.     

Net mineral requirements for the growth and maintenance of Somali lambs

Minerals are limiting factors in animal production, and the knowledge of mineral requirements for livestock is crucial to the success of a commercial enterprise. Hair sheep may have different mineral requirements than those presents by the international committees. A study was carried to evaluate the net calcium (Ca), phosphorus (P), magnesium (Mg), sodium (Na), potassium (K), zinc (Zn), iron (Fe), manganese (Mn) and copper (Cu) requirements for the growth and maintenance of Brazilian Somali lambs. A total of 48 hair lambs (13.5±1.8 kg) aged 60±15 days were allocated to individual pens. Eight animals were slaughtered at the beginning of the experiment to serve as a reference group to estimate initial empty BW (EBW) and initial body composition. The remaining lambs (n=40) were assigned to a completely randomized design with eight replications in five levels of metabolizable energy (ME; 4.93, 8.65, 9.41, 10.12 and 11.24 MJ/kg DM). When the lambs of a given treatment reached an average BW of 28 kg, they were slaughtered. Initial body composition was used to calculate the retention of minerals. Mineral body composition was fit using a logarithmic equation in the form of a nonlinear model. The maintenance requirements were estimated from regressions of mineral retention in the empty body on mineral intake. The body mineral concentration decreased in lambs with a BW ranging from 15 to 30 kg. The net mineral requirements (100 g/day of average daily gain (ADG)) decreased from 0.52 to 0.51 g for Ca, 0.28 to 0.23 g for P, 0.02 to 0.02 g for Mg, 0.09 to 0.08 g for Na, 0.11 to 0.09 g for K, 1.30 to 1.08 mg for Zn, 3.77 to 3.22 mg for Fe, 0.08 to 0.06 mg for Mn and 0.09 to 0.08 mg for Cu when BW increased from 15 to 30 kg. The daily net requirements for maintenance per kilogram of BW were 30.13 mg of Ca, 27.58 mg of P, 1.26 mg of Mg, 4.12 mg of Na, 8.11 mg of K, 0.133 mg of Zn, 0.271 mg of Fe, 0.002 mg of Mn and 0.014 mg of Cu. The results of this study indicate that the net mineral requirements for weight gain and maintenance in Brazilian Somali lambs are different than the values that are commonly recommended by the main evaluation systems for feed and nutritional requirements for sheep. These results for the nutritional requirements of minerals may help to optimize mineral supply for hair sheep.     

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.     

Purification of 4S-limonene synthase, a monoterpene cyclase from the glandular trichomes of peppermint (Mentha x piperita) and spearmint (Mentha spicata).

The p-menthane monoterpenes of the Mentha species are biosynthesized from geranyl pyrophosphate via the monocyclic olefin 4S-limonene. A monoterpene cyclase was isolated from both Mentha x piperita (peppermint) and Mentha spicata (spearmint) that catalyzes the cyclization of geranyl pyrophosphate to 4S-limonene. This enzyme, 4S-limonene synthase, was purified to apparent homogeneity by dye ligand, anion exchange, and hydrophobic interaction chromatography. Since the monoterpenes of Mentha are synthesized and secreted in modified epidermal hairs called glandular trichomes, an extract of isolated glandular trichome cells was used as the source of this enzyme. A combination of gel permeation chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that purified 4S-limonene synthase had a native molecular weight of 56,000 and was monomeric. The principal product of the enzyme was enantiomerically pure (-)-4S-limonene, and a catalytic constant of 0.3/s was determined. The basic properties of 4S-limonene synthase from both M. x piperita and M. spicata are identical and, in general, are similar to those of other monoterpene, sesquiterpene, and diterpene cyclases isolated from microorganisms and higher plants.