In Vitro Gynogenesis in Leek (Allium ampeloprasum L.).

Leek (A. ampeloprasum L.) is an economically important vegetable crop from Alliaceae family. It is a non-bulb forming biennial species grown for its pseudostem and leaves. Leek is a tetraploid with one of the largest genomes known among cultivated plant species. It has enormous economic importance all around the world for many purposes such as vegetable, medicinal herb, and food seasoning. Production and consumption of leek is in rise all around the world and breeders are trying to develop new F1 hybrid varieties with desired agronomical traits. Although self-compatible, leek shows high tendency toward outcrossing and display severe inbreeding depression when selfed with its own pollen. Therefore, inbred development through classical breeding techniques is very difficult in this crop. Traditional leek genotypes are highly heterozygous, open pollinated varieties. There is a high demand for F1 hybrid varieties with resistance to biotic and abiotic stresses and high-quality plants. Our group is trying to incorporate gynogenesis-based doubled haploid technology to leek improvement programs. Over the years, many experiments were carried out to determine the gynogenic potential of donor leek genotypes of different genetic backgrounds in different induction media. Here, we report a protocol allowing production of green gynogenic leek plants via single step culture of unopened flower buds. Ploidy levels of gynogenic regenerants are determined by flow cytometry analysis. A majority of the gynogenic leek regenerants produced survived well in vivo.

Selenium and Sulfur to Produce Allium Functional Crops.

Selenium is an element that must be considered in the nutrition of certain crops since its use allows the obtaining of biofortified crops with a positive impact on human health. The objective of this review is to present the information on the use of Se and S in the cultivation of plants of the genus Allium. The main proposal is to use Allium as specialist plants for biofortification with Se and S, considering the natural ability to accumulate both elements in different phytochemicals, which promotes the functional value of Allium. In spite of this, in the agricultural production of these species, the addition of sulfur is not realized to obtain functional foods and plants more resistant; it is only sought to cover the necessary requirements for growth. On the other hand, selenium does not appear in the agronomic management plans of most of the producers. Including S and Se fertilization as part of agronomic management can substantially improve Allium crop production. Allium species may be suitable to carry out biofortification with Se; this practice can be combined with the intensive use of S to obtain crops with higher production and sensory, nutritional, and functional quality.

A mutagenicity and cytotoxicity study of limonium effusum aqueous extracts by Allium, Ames and MTT tests.

Nowadays plants or plant extracts have become very important for alternative medicine. Plants and their extracts have many therapeutical advantages but some of them are potentially toxic, mutagenic, carcinogenic and teratogenic. Root, stem and leafparts of Limonium effusum were used in this study and this species is an endemic species for Turkey. Mutagenic and cytotoxic effects of root, stem and leaf aqueous extracts were observed with Allium, Ames and MTT tests. Allium root growth inhibition test and mitotic index studies showed that aqueous extracts have dose-dependent toxic effects. Chromosome aberration studies indicated that especially sticky chromosome, anaphase-telophase disorder and laggard chromosome anomalies were highly observed. Ames test performed with Limonium effusum root aqueous extracts, showed weak mutagenic effects in Salmonella typhimurium TA98 strain with S9. MTT test based on mitochondrial activity indicated that most of the aqueous extracts have cytotoxic effects. This study aimed to determine the possible mutagenic and cytotoxic effects of L. effusum aqueous extracts by using bacterial, plant and mammalian cells. This research showed that some low concentrations of the L. effusum extracts have inhibited cytotoxic effects but high concentrations have induced cytotoxicity. On the other hand only a weak mutagenic activity was identified by Ames test with TA98 S9(+).

Shading decreases plant carbon preferential allocation towards the most beneficial mycorrhizal mutualist.

Preferential allocation towards the most beneficial mutualist could maintain mycorrhizal mutualism. Context dependence of preferential allocation could then determine environmental patterns in abundance of mycorrhizal mutualists. We assessed the preferential allocation of carbon (C) and differential phosphorus (P) uptake across four light treatments between the host plant Allium vineale and two arbuscular mycorrhizal (AM) fungi within a split-root system. The ratios of C allocation and P uptake between the beneficial and nonbeneficial AM fungi were measured using isotopic labelling. Allium vineale preferentially allocated more C towards roots infected with the most beneficial AM fungus in high light and, in return, received more P from the beneficial fungus. Preferential allocation declined with shading, as A. vineale allocated 25% of labelled C to roots infected with beneficial AM fungi in high light, but only 15% with shading, a similar percentage to that allocated to roots infected with nonbeneficial fungi regardless of shading. Our findings demonstrate that plant preferential allocation towards the most beneficial mycorrhizal mutualist depends upon above-ground resources, suggesting that the abundance of beneficial mycorrhizal fungi will increase with amount of above-ground resources, with implications for mycorrhizal mediation of plant productivity with anthropogenic change.

Phytotoxicity of secondary metabolites isolated from Flourensia oolepis S.F.Blake.

The aim of this study was to isolate the active principles of Flourensia oolepis S.F.Blake (Asteraceae), which completely inhibited the germination of Raphanus sativus seeds at 10 mg/ml. Flavanone pinocembrin and sesquiterpene ilicol, were isolated by bioassay-guided fractionation. They were active both against monocot and dicot seeds. Pinocembrin was the most active compound, with an IC50 (germination) value of 0.24, 3.40, 3.28, and 3.55 mM against Panicum miliaceum, Avena sativa, Lactuca sativa, and R. sativus, respectively; ilicol, however, exhibited IC50 (germination) values of 0.67, 2.73, 5.25, and 9.66 mM for the same species, respectively. Pinocembrin and ilicol inhibited root growth and showed IC50 (root growth) values of 0.199, 14.68, 8.05, 7.69 mM, and 1.22, 2.90, 7.35, 8.07 mM, against P. miliaceum, A. sativa, L. sativa, and R. sativus, respectively. Pinocembrin and ilicol reduced Allium cepa cell division without chromosome aberrations.

Phytotoxicity of lignanamides isolated from the seeds of Hyoscyamus niger.

Bioassay-guided fractionation of phytotoxic extracts prepared from the seeds of Hyoscyamus niger led to the isolation of three new lignanamides (1-3), along with six known lignanamides (4-9). The structures of the new compounds were determined by spectroscopic methods, including 1D and 2D nuclear magnetic resonance techniques, and high-resolution electrospray ionization mass spectrometry. The bioactivity analysis of the isolated compounds showed that compound 3 exhibited significant inhibition on the germination and radical elongation of Allium fistulosum at 10(-4) M concentration.

Variations between diploids and tetraploids of Allium przewalskianum, an important vegetable and/or condiment in the Himalayas.

Allium przewalskianum, a wild onion species growing at altitudes ranging from 1800 to 4500 m, has long been commonly used as an important vegetable and/or condiment by Tibetans, Indians, and Nepalese in the highlands of the Himalayas and adjacent regions. This species comprises both diploids and tetraploids. In this study, we examined the nutritional content and biomass accumulation profiles of two cytotypes, collected from 29 sites, with different altitudinal origins but cultivated in a common garden. On an average, this species has superior qualities in the minerals and amino acids compared to other edible congeners. When compared with the diploids, the tetraploids grew faster and accumulated more biomass; in addition, the tetraploids had higher values of moisture and energy, higher contents of cystine and phosphorus, but lower fiber levels. Moreover, the tetraploids from the higher altitudes had greater biomasses than the other tetraploids, in addition to having increased levels of proteins, fats, and the minerals Mg, Fe, Mn, and Cu. These results illustrate the large variations in nutritional efficacy and growth within this single morphological species, and provide critical information for its effective consumption in the future.

A comparative study of the antimicrobial and phytochemical properties between outdoor grown and micropropagated Tulbaghia violacea Harv. plants.

AIM OF THE STUDY: The study aimed to compare the antimicrobial and phytochemical properties of in vitro cultured and outdoor grown Tulbaghia violacea plants in the quest to validate the use of micropropagated plants as alternatives to outdoor grown plants in traditional medicine. Tulbaghia violacea is used extensively in South African traditional medicine for HIV/AIDS patients and in the treatment of gastrointestinal ailments, asthma, fever and tuberculosis. MATERIALS AND METHODS: Extracts of micropropagated and outdoor grown Tulbaghia violacea plants were evaluated for their antibacterial and antifungal activities against Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and a fungus Candida albicans using microdilution methods. Saponins and phenolic compounds including condensed tannins, gallotannins and flavonoids were quantitatively determined using spectrophotometric methods. A qualitative test for saponins was also carried out. RESULTS: The petroleum ether (PE) extracts of micropropagated plants and dichloromethane (DCM) extracts of outdoor grown plants showed good antibacterial activity, each against two bacterial test strains. PE extracts of micropropagated plants showed the best antibacterial activity with a minimum inhibitory concentration (MIC) of 0.39 mg/ml against Bacillus subtilis. Good MIC (<1mg/ml) and minimum fungicidal concentration (MFC) values of 0.78 mg/ml were only obtained in DCM extracts of outdoor grown plants. MIC and MFC values for water and ethanol extracts of both micropropagated and outdoor grown plants were similar and in the range 3.125-12.5mg/ml. Total phenolics, gallotannins, flavonoids and saponins were significantly higher in micropropagated plants than in outdoor grown ones. In all cases, the amounts of phytochemical compounds in micropropagated plants were more than twice that of outdoor grown plants except for condensed tannins. CONCLUSION: The results form a good basis for the use of Tulbaghia violacea micropropagated plants as a complement to the outdoor grown plants in traditional medicine.

Insecticidal, mutagenic and genotoxic evaluation of annonaceous acetogenins.

Annonaceous acetogenins represent a class of bioactive compounds whose primary mode of action is the inhibition of NADH-ubiquinone oxidoreductase (Mitochondrial Complex I). Given the potential pesticidal use of these compounds, we evaluated the effects of seven acetogenins: squamocin (1), molvizarin (2), itrabin (3), almuñequin (4), cherimolin-1 (5), cherimolin-2 (6), and tucumanin (7) isolated from Annona cherimolia Mill. against Ceratitis capitata Wiedemann (Tephritidae). These acetogenins did not display insecticidal action at 250 microg of treatment per g of adult diet. However, the oviposition capacity of C. capitata females was significantly altered by some of the acetogenins at this concentration. The most potent compounds were itrabin, molvizarin and squamocin. Moreover, significant differences were detected in the preference of oviposition sites when itrabin and squamocin were spread on the surface of artificial fruits at doses of 30 microg/cm2. Additionally, we investigated the mutagenic effects displayed by itrabin, as well as the phytotoxic and genotoxic action of squamocin and itrabin. Both compounds displayed slight phytotoxic and genotoxic effects on roots of Allium cepa at 2.5 microg/mL though no mutagenic effects were detected at 0.25, 0.5 and 2.5 microg/mL on Salmonella typhimurium strains TA98 and TA100.

Investigation of cytotoxic and genotoxic effects of Ecballium elaterium juice based on Allium test.

The genus Ecballium only comprises the Ecballium elaterium (EE) (L.) A.Rich species which is a wild medicinal plant found in the Mediterranean region. EE fruit juice is widely used in Turkish folk medicine for the relief of sinusitis and for several illnesses. Up to date, there has been no report on the genotoxicity of EE fruit juice. Thus, the aim of this study was to investigate the potential genotoxic effects of EE fruit juice using the Allium test system. Allium cepa (A. cepa) bulbs were treated with four concentrations (10 ml/L, 20 ml/L, 50 ml/L and undiluted) of EE fruit juice for 72 h and tap water (pH 7.3) was used as a control. The results showed significant dose-dependent (P < 0.05) inhibition of root growth and mitodepressive effects on cell division in A. cepa root tip cells after the EE fruit juice treatments. Also, EE fruit juice significantly increased the dose-dependent frequency of chromosome aberrations (breaks, stickiness and pole deviations) in root tip cells and micronucleus formations. There was no dividing cell in the undiluted EE fruit juice treated group, but there were pyknotic/apoptotic cells with varying frequency.

Are there benefits of simultaneous root colonization by different arbuscular mycorrhizal fungi?

Arbuscular mycorrhizal fungal (AMF) communities were established in pots using fungal isolates from a single field in Switzerland. It was tested whether multispecies mixtures provided more phosphorus and supported greater plant growth than single AMF species. Two host plants, medic (Medicago truncatula) and leek (Allium porrum), were inoculated with three AMF species (Glomus mosseae, G. claroideum and G. intraradices), either separately or in mixtures. The composition of the AMF communities in the roots was assessed using real-time PCR to determine the copy number of large ribosomal subunit genes. Fungal communities in the roots were usually dominated by one AMF species (G. mosseae). The composition of the communities depended on both plant identity and the time of harvest. Leek colonized by a mixture of G. claroideum and G. intraradices acquired more P than with either of the two AMF separately. Direct evidence is provided for functional complementarity among species within the AMF community colonizing a single root system. Competition among the species poses a major challenge in interpreting experiments with mixed inoculations, but this is greatly facilitated by use of real-time PCR.

Influence of nitrogen fertilizers on yield and antifungal bioactivity of Tulbaghia violacea L.

Tulbaghia is known to have antifungal properties that can be used in the treatment of both human and plant pathogens and is used in traditional medicine in South Africa. Increasing demands for plant material makes it necessary to cultivate this species on a large scale. Unfortunately, cultivation can lead to a reduction in the biological activity of plants making them unsuitable for use. In light of the lack of knowledge regarding the agronomic requirements of this plant, the aim of this study was to determine the effect of several rates and two forms of nitrogenous fertilizer on the yield and biological activity of Tulbaghia violacea. Plants were cultivated in sand while the basic fertilization used was the same as that of garlic (20 kg P ha(-1), 75 kg K ha(-1)), a plant from the same family, containing similar active ingredients. Nitrogen was applied once at the beginning of the trial at rates of 30, 60, 120, and 180 kg ha(-1) in the form of either nitrate or ammonium. Vegetative growth was quantified in terms of number of leaves and leaf area as well as root and leaf dry mass, while harvested material was tested for antifungal activity. The results indicated that compared with the untreated control, increasing N-rates in both the nitrate and ammonium forms increased leaf number and leaf area as well as both root and leaf dry mass. However, at a rate above 60 kg ha(-1), and especially at 180 kg ha(-1), the nitrate form stimulated growth more markedly than the ammonium form, whereas antifungal activity decreased sharply and almost linearly as the application rate was increased. Although growth was not stimulated to the same extent by the ammonium form, it increased the in-vitro antifungal activity at different levels during different times of the growing season. From an ornamental perspective, nitrate is the preferred nitrogenous form but, from a bioactivity perspective, ammonium is recommended.

Chemical characterization of Allium ursinum L. depending on harvesting time.

Sulfur-containing compounds of ramson (Allium ursinum L.) are responsible for its traditional use in terms of culinary and medicinal purposes. Leaves and bulbs were investigated for their contents of cysteine sulfoxides (volatile precursors) as well as volatile compounds released from minced plant material. Plants were analyzed during the whole vegetation period, focused on the months from March to June. Additionally, within the dormancy period bulbs were analyzed again and alliinase activity was determined. The pattern of volatile compounds was analyzed both by SPME/GC-MS and by SDE/GC-MS. Compared to each other, SDE exhibited a wider spectrum of detectable volatile compounds. The quality and quantity of volatiles significantly depended on the time of harvest. The highest amounts of volatile precursors can be gained in March and April, shortly before flowering time (up to 0.4% of total cysteine sulfoxides). The main cysteine sulfoxides were alliin and isoalliin. It has been found that alliinase of A. ursinum exhibited properties similar to those of alliinase of garlic (Allium sativum L.), but differing in terms of substrate specificity.

Improving seedling vigour of indigenous medicinal plants with smoke.

The application of smoke and aqueous smoke solutions stimulates seed germination in a number of plant species. This study highlights the effects of aerosol smoke and smoke solutions on the germination and seedling vigour of three South African indigenous medicinal plants Albuca pachychlamys, Merwilla natalensis and Tulbaghia violacea. The vigour index of one-week-old seedlings of all three species examined was increased with the application of dry smoke and smoke extract dilutions, as compared to control treatments. Seedlings of A. pachychlamys germinated with smoke solutions showed a significant (p0.05) gain in bulb and leaf mass (27.9 and 197.6 mg respectively) compared to untreated seedlings (9.9 and 124.7 mg respectively) when grown in vitro for 75 days. The leaf mass of smoke solution-treated seedlings of T. violacea was significantly (p0.05) higher (120.4 mg) than that of untreated seedlings (47.6 mg). Subsequently, the height of seedlings in both species was also significantly (p0.05) greater. Seedlings germinated in water and then transferred to smoke solutions (1:2000) showed enhancement of some of the growth parameters studied. Albuca pachychlamys and T. violacea seeds exposed to aerosol smoke exhibited higher seedling survival percentages than from non-smoked seeds, while no significant effect was observed for M. natalensis seedlings. This investigation shows that the application of smoke technology can be adopted to produce high vigour seedlings.

Tumorigenesis, metabolism, speciation, bioavailability, and tissue deposition of selenium in selenium-enriched ramps (Allium tricoccum).

Ramps (Allium tricoccum) were grown either in a mixture of vermiculite and peat moss or hydroponically with various concentrations of selenium as sodium selenate. The concentrations used were from 30 to 300 mg of selenium/kg of vermiculite-peat moss or from 10 to 120 mg/L in the hydroponic solutions. Levels as high as 784 mg of selenium/kg were obtained in the ramp bulbs when grown with high levels of selenium in the vermiculite-peat moss, and up to 600 mg of selenium/kg was obtained hydroponically. The predominant form of selenium in the ramp bulbs at all concentrations of selenium was Se-methylselenocysteine, with lower amounts of selenate, Se-cystathionine, and glutamyl-Se-methylselenocysteine. There was a approximately 43% reduction in chemically induced mammary tumors when rats were fed a diet with Se-enriched ramps. Dietary Se-enriched ramps for rats did not result in excessive tissue selenium accumulation or undesirable side effects. Bioavailability studies with rats indicated that selenium in ramps was 15-28% more available for regeneration of glutathione peroxidase activity than inorganic selenium as selenite. Therefore, Se-enriched ramps appear to have potential for the reduction of cancer in humans.

Growing alliums and brassicas in selenium-enriched soils increases their anticarcinogenic potentials.

The concentrating of essential minerals, vitamins, and bioactive phytochemicals into human foodstuffs is of vital importance in our rapidly expanding world. Selenium is an essential micronutrient which is currently receiving much publicity for its anticarcinogenic potentials. Unfortunately, this mineral is deficient in most soils worldwide, and as a result most geographical food chains contain highly inadequate amounts of selenium. Scientific evidence in now available which shows that common vegetable members of the allium genus, especially garlic, are seleniferous in that they readily uptake inorganic selenium from the soil and incorporate it into bioactive organic chemicals. The brassicas have also been found to be highly seleniferous in nature and to produce various seleno amino acids and potentially bioactive organic selenium-containing phytochemicals. Upon consumption by humans, these selenium phytochemicals, derived from both plant genus, show anticarcinogenic potentials. Due to the high concentrations of natural phytochemicals, and the additional assimilation of selenium, the commercial or small scale production of selenium-enriched brassicas and garlic is an excellent way of introducing anticarcinogenic phytochemicals into the human diet.