Mojdeh Asadi; Javad Hadian; Samad Nejad Ebrahimi; Ghasem Karimzadeh
Abstract
Introduction: The genus Arnica L. comprises of 32 species predominantly confined to the boreal and montane region of the northern hemisphere. Arnica species are rhizomatous perennial herbs belonging to the daisy family, with simple or branched stems bearing opposite leaves, and large, single or cymose ...
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Introduction: The genus Arnica L. comprises of 32 species predominantly confined to the boreal and montane region of the northern hemisphere. Arnica species are rhizomatous perennial herbs belonging to the daisy family, with simple or branched stems bearing opposite leaves, and large, single or cymose heads of yellow flowers. A. chamissonis is distributed over North America from Alaska to New Mexico, and due to its low ecological demand it is easier to cultivate than A. montana. Arnica is a source of sesquiterpene lactones, flavonoids, essential oils, terpenoids, and phenolic acids and exhibits antiseptic, anti-inflammatory, antiradical, antibacterial, anti-sclerotic, antifungal, and antioxidant activities. The flower heads and other parts of the plant of two of the species have been used therapeutically: A. montana and A. chamissonis, both of them containing sesquiterpene lactones as pharmacologically active compounds. Arnica species are used in as many as 300 drug preparations in Europe and about 20 products in Canada. Moreover, A. chamissonisis a good source of bioactive compounds a valuable source of herbal raw material and a pharmaceutical substitute for the endangered mountain arnica. A. chamissonis extracts exhibit potent anti-inflammatory and anti-radical activity and possesses high antioxidant abilities that might be helpful in preventing or slowing the progress of free radical-dependent diseases. Low pH soil is one of the principal ecological requirements of Arnica. Good plant growth and flower yield was achieved on acid soils with a pH of 6.8 and below. After nitrogen, phosphorus and potassium, sulfur is considered as fourth major element in most crops. Rate of sulfur oxidation in soils vary and depend on population Thiobacillus bacteria in soil, particle size and environmental conditions.
Materials and Methods: This study was conducted as a complete randomized block design with three replications and four treatments including different levels of bentonite sulfur (0, 250, 500 and 750 kg/ha) combined with the bacterium Thibacillus thiooxidans. Traits such as height and width of plants, number of flowers on each plant, inflorescence diameter, fresh and dry yield of flowers in each plant per square meter and total phenols, total flavonoids, rutin, luteolin and apigenin content in flowers were measured. High-Performance Liquid Chromatography method (HPLC) was used to separate rutin, luteolin and apigenin. HPLC grade methanol and distilled water, each with 0.02% added TFT were used as solvents.
Results and Discussion: The results showed that A. chamissonis is compatible with Tehran’s climate and it can be cultivated in regions with a similar climate. The use of elemental sulfur has a significant role in reducing soil pH and soil pH decreased more rapidly with increasing sulfur. Thiobacillus bacteria by oxidation of sulfur produced some to sulfuric acid and at low buffered properties can considerably reduce pH. Different levels of sulfur fertilizer had a significant effect (P ≤0.01) on soil acidity (pH), electrical conductivity, and sulfur, iron, zinc and manganese levels. The use of 750 kg sulfur bentonite with Thiobacillus caused to reduce soil acidity by about 0.9 units, and it increased the electrical conductivity of the soil to 7.33 dS/m. The amount of soil sulfate as a result of oxidation of sulfur fertilizers increased linearly. The highest amounts of iron, zinc and manganese were measured in the treatment of 750 kg sulfur as 3.38, 3.84 and 27.94 mg /kg, respectively. In this study, supplying 250 kg of sulfur plus bio-sulfur caused to improve the morphological traits and the highest flower yield (20.67 g / plant) compared to control (3.66 g / plant). Also, it increased the amount of sulfur and micro-elements in the soil, but in the treatment of 500 and 750 kg sulfur per hectare due to increased electrical conductivity and soil salinity, the growth of Chamisso Arnica decreased. Therefore, these levels of sulfur are not suitable for this plant. Sulfur Bentonite is oxidized by Thiobacillus bacteria and other microorganisms and sulfuric acid is produced. Reducing acidity and, as a result, the release of stabilized and insoluble nutrients can increase their uptake capacity and improve plant growth and development. Due to the increased salinity and osmotic imbalance in the soil by the treatments of 500 and 750 g /kg sulfur, vegetation growth will be incomplete and plants quickly enter to the reproductive phase and produce fewer and smaller flowers. The highest content of rutin was measured in plants grown in soil treated with 750 Kg/ha sulfur while soil treatment with 500 Kg/ha sulfur resulted in highest content of luteolin and apigenin, both showing significant difference to the control.
Conclusion: Results of this study indicated that the Chamisso Arnica showed adaptability to the planting location and it can be mass-cultivated under similar conditions.
Shadab Faramarzi; Abbas Yadollahi; Ghasem Karimzadeh
Abstract
Introduction: Apple (Malus Miller) belongs to Rosacease family and the subfamily of Pomoideae. This fruit is at first place among fruits of temperate zones. The cultivated apple (Malus × domestica Borkh.) is a complex hybrid of the apple species. Chromosomal basis of this subfamily are x = 17 and the ...
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Introduction: Apple (Malus Miller) belongs to Rosacease family and the subfamily of Pomoideae. This fruit is at first place among fruits of temperate zones. The cultivated apple (Malus × domestica Borkh.) is a complex hybrid of the apple species. Chromosomal basis of this subfamily are x = 17 and the ploidy levels have been reported for diploid (2n=2x = 34), triploid (2n =3x= 51) and tetraploid (2n= 4x= 68). Since Iran is close to the apple diversity region (Central Asia), it has a good variation of apple varieties. Despite the high levels of variation in apple cultivars and species in Iran, there is not still a database of genome size. Classification of plants according to their genome size, especially at lower taxonomic levels is important for breeders. Over the past years, several methods for estimation of nuclear DNA content (genome size) was common, but recently, the use of flow cytometry (FCM) has been increasingly used. Flow cytometry is the best method to estimate DNA c-value and ploidy levels in apples. In this study, DNA c-Value and ploidy level of Iranian apple varieties has been estimated by flow cytometry and propidum iodide staining.
Materials and Methods: Fully expanded young leaves of all apple varieties were collected in the summer 2013. Nuclear extraction was performed using Partec kit as following: 1 cm2 apple leaf and 1 cm2 parsley leaf (as internal standard) were co-chopped with razor blade after adding 500 µl of nuclear extraction buffer. Then, the extract was filtered by two kind of filters (50 and 30 um). One ml of staining buffer, 4 µl of RNAase and 4 µl ofpropidium iodide was added for 15 min at room temperature. Finally, nuclei were counted using flow cytometry (BD FACSCanto II, USA) at Tarbiat Modarres University. The genome size was estimated according to bellow formula:
DNA 2C-value sample =
Also, given the high levels of phenolic compounds in apples, treating with PVP and PVP 1% were performed to evaluate the effect of phenolic compounds on estimation of genome size. Finally, Histogram analysis and DNA c-value estimation were done with Partec Flow Max software. The difference between means was obtained by SAS software ver. 9.2 and LSD tests.
Results and Discussion: The results showed that genome size obtained from Partec Flow Max software and ranged from 1.57 pg for ‘Golab- Bastam’ to 1.73 pg for ‘Golab- Kermanshah’. Histogram analysis was demonstrated that all studied cultivars are diploid. The average genome size in this study was 1.62 pg. Research conducted on foreign apple varieties have showed that the genome size of diploid species from was obtained 1.45 for M. fusca to1.68 pg for M. ransitoria. The genome size for triploid species was ranged from 2.37 to 2.57 pg. In this study, genome size was calculated in terms of mega base pairs and was different from 748 Mbp in ‘Golab- Bastam’ to 846 Mbp in ‘Golab- Kermanshah’. Thus, the size of the genome was closed to M. ransitoria (1.68 pg). This species is native to China, which is a Crab apple and used as an ornamental tree. It has been reported that Iranian apple are M. domestica Borkh. In another study, genome size was identified in the range from1.245 pg for diploid species of M. tschonoskii to 1.653 pg for M. florentina. M. florentina species is native to Balkans and Italy, that is an ornamental tree and its genome size is close to M. domestica Borkh. (1.653 pg).
Conclusion: Classification of plants according to their genome size seems to be important, especially at lower taxonomic. Genome size, even in very close species can also be different, for example, northern corn with more heterochromatin has larger genomes than those who are located in south (less heterochromatin). This study appears the variation of DNA 2C-value in Golab cultivars, even though Golab cultivars are known clones with low genetic diversity. Therefore, it is likely that Iranian apple varieties, with the same ploidy level, have been had difference in genome size. There are various ploidy level in apple, including diploid (2n = 34), triploid (2n = 51), tetraploid (2n = 68) and hexaploid (2n = 102). Thus, it is expected that current apple M. × domestica Borkh., have been contributed some several species such as M. prunifolia (Willd.) Borkh., M. baccata (L.) Borkh., M. sieboldii (Regel) Rehder, M. sylvestris, ،M. orientalis Uglitzk and M. sieversii.
