Esmaeil Chamani; Zahra Eftekhari; Alireza Ghanbari; Hamid Reza Heydari; Mousa Arshad
Abstract
Introduction: Fritillaria imperialis L. is an ornamental and medicinal plant native to mountainous regions of Iran. This plant genetic resource is in danger of extinction, Because of grazing livestock and pest outbreaks. Therefore, micro propagation of Fritillaria through in vitro regeneration is essential ...
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Introduction: Fritillaria imperialis L. is an ornamental and medicinal plant native to mountainous regions of Iran. This plant genetic resource is in danger of extinction, Because of grazing livestock and pest outbreaks. Therefore, micro propagation of Fritillaria through in vitro regeneration is essential for conservation and commercial production. Thymol and Carvacrol are one of the main essential oil compounds in family Lamiaceae.
Material and Methods: Fritillariaimperialis L. bulbs in dormancy stage obtained from mountainous regions of Lorestan in Iran and were placed in cold room at +4 °C for 4-6 weeks. Then, Bulbs were surface-sterilized with 70% ethanol for 45 s followed by immersion in 5% (v/v) NaOCl solution for 20 min with gentle agitation, and then rinsed three times in sterile double distilled water. Present study was conducted in two separate experiments. In first experiment, effect of different concentration of Thymol and Carvacrol and in second experiment, different concentration of NAA and BA on in vitro characteristics of Fritillaria was evaluated. Explants (1× 1 cm) prepared from the lower third of scales with basal plate and were placed in MS basal medium supplemented with different concentrations of Thymol (50, 100, 150 and 300 ppm), Carvacrol (10, 100, 500 and 100 ppm), BA (1, 2 and 4 mg/l) and NAA (1, 2 and 4 mg/l).All cultures were incubated in a growth chamber at 24±2°C, and a photosynthetic photon flux of 40-60 μmol m–2 s–1 was provided by cool white fluorescent lamps with a 16-h photoperiod. This experiment wascarried out in completely randomized designs with fivereplications.
Results and Discussion: Analysis of variance showed that Thymol and Carvacrol were not effective on number of new bulblets but had significant effects on bulb diameter, number and length or roots, number and length leaves and callus induction and diameter of callus obtained from scales (P< 0.05). The highest rate (3 bulblets) of bulblets formation was obtained fromMS medium supplemented with 50 ppm Thymol that showed significantly difference from other treatments. Medium containing 10 ppm Carvacrol gave the highest Bulblet formation (2.5 bulblets) between Carvacrol treatments. Investigation of rooting was done by assessment of the number and length of roots. Mean comparison of the effect of cultivar type on root number showed that the largest number of roots per explant was obtained fromMS medium containing 50 ppm Thymol. Lowest number of roots observed in mediums supplemented with 300 ppm Thymol and 100 ppm Carvacrol. The best medium for increasing the root length per explant (10.90 cm) was MS medium supplemented with 100 ppm Carvacrol, while the least increasing in root length per explant observed from culture mediums contained 300 ppm Thymol and 100 ppm Carvacrol. Also, the largest number of leave formation obtained from culture medium supplemented with 50 ppm Thymol that significantly higher than other treatments. Statistical analysis (ANOVA) of the data showed that high frequency callus induction and formation occurred in MS mediums contained 50, 100 and 150 ppm Thymol and 10 ppm Carvacrol and culture mediums supplemented with 300 ppm Thymol and 1000 ppm Carvacrol showed least callus induction. In contrast, largest callus diameter observed in culture mediums supplemented with 300 ppm Thymol and 500, 100 ppm Carvacrol.
Statistical analysis of results showed that different concentrations of BA and NAA had significant effects on bulblets number and bulblets diameter (P
Esmaeil Chamani; Marzyeh Ghamari; Mahdi Mohoboldini; Alireza Ghanbari; Hamidreza Heydari
Abstract
Introduction: Crown imperial (Fritillariaimperialis L.) is an ornamental and medicinal plant native to mountainous regions of Iran. This plant genetic resources is in danger of extinction, because of grazing livestock and pest outbreaks. However, due to slow reproduction in natural conditions and traditional ...
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Introduction: Crown imperial (Fritillariaimperialis L.) is an ornamental and medicinal plant native to mountainous regions of Iran. This plant genetic resources is in danger of extinction, because of grazing livestock and pest outbreaks. However, due to slow reproduction in natural conditions and traditional multiplication methods such as scaling and Bulb division, many species of this genus are endangered. Using of biotechnology, namely in vitro plant propagation, is a solution to the problems of reproduction of rare and endangered plant species with difficult propagation and mass production of valuable genotypes. Therefore, micropropagation of F. imperialis through in vitro regeneration is essential for conservation and commercial production.
Material and Methods: The bulbs of F. imperialis in dormancy stage obtained from Ilam mountainous regions in Iran and theywere placed in wet vermiculite at 4 °C for 4-6 weeks. Then, Bulbs were surface-sterilized with 70% ethanol for 60s followed by immersion in 5% (v/v) NaOCl solution for 20min with gentle agitation, and they rinsed three times in sterile double distilled water. Explants prepared from the lower third of scales with basal plate and were placed in MS basal medium supplemented with different concentrations of NAA and 2,4-D for callus induction. Test tubes with bulb segments were maintained within 25±2°C in growth chamber at 16 hours light period by the illumination from white florescent tube light and 8 hours dark. After two months callus were transferred to MS basal medium without PGRs. Then, callus excised to 0.5 cm pieces and were transferred to MS basal medium supplemented with NAA in 0, 0.3 and 1 mg/l concentration.Three types of cytokinins with different concentrations were arranged in three seperated experiments. Thefirst experiment medium contained NAA with BA (0, 0.3, 0.5 and 1 mg/l), the second experiment NAA combined with 0, 0.1, 0.3 and 0.5 mg/l TDZ and the third experiment MS basal medium included NAA with Kin (0, 0.5, 1 and 1.5 mg/l). After three months, percentage of callogenesis, diameter of calli, percentage of regeneration, number of leaves and roots and length of leaves and roots were measured. This experiment were carried out in completely randomized design with 4 replications.
Results and Discussion: In the first experiment application of NAA and BA on in-vitro multiplication of F. imperialis were evaluated. Highest callogenesis and formation (100 %) was observed in mediums contained 0.3 mg/l NAA + 1 mg/l BA, 0.6 mg/l NAA + (0.3, 0.5 and 1 mg/l) BA. Also, callogenesis was obtained in medium contained 0.5 mg/l BA without NAA. This result showed that only in medium supplemented with 1 mg/l BA provided highest (100%) callogenesis, when NAA concentrations were low. However, high levels of NAA (0.6 mg/l) in all concentrations of BA were obtained maximum callogenesis. We concluded that NAA is essential for callogenesis and enhancing its levels can increase callogenesis. Also, application of low levels of BA (0.4 µM) in callogenesis mediums of Cynodon dactylon contained Auxins resulted in increment of embryogenetic calli formation. In the other hand, presence of BA is essential for plantlet regeneration, however NAA is not necessary. Plantlet regeneration was obtained in PGRs free medium. Statistical analysis of results showed that different concentrations of BA and NAA had significant effects on percentage of callogenesis, diameter of calli, percentage of regeneration, length of leaves and roots (P
E. Chamani; M. Bonyadi; A. Ghanbari
Abstract
Introduction: Vinca flower (Catharanthus roseus L.) is one of the most important medicinal plants of Apocynaceae (31, 27). Tropical plant native to a height of 30 to 35 centimeters (9) and a perennial shrub which is grown in cold areas for one year (27).One of the plants in the world today as a medicinal ...
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Introduction: Vinca flower (Catharanthus roseus L.) is one of the most important medicinal plants of Apocynaceae (31, 27). Tropical plant native to a height of 30 to 35 centimeters (9) and a perennial shrub which is grown in cold areas for one year (27).One of the plants in the world today as a medicinal plant used the periwinkle plant. Among the 130 indole – terpenoids alkaloids which have been identified in the plant periwinkle vinca alkaloids vincristine and vinblastin are the most important component is used to treat a variety of cancers. Including therapies that are used for a variety of cancer, chemotherapy to help Vinca alkaloids collection (including vincristine and…). Vinblastin as effective member of this category, due to the low percentage of venom and effects at very low doses, is widely used today. These materials are generally formed as inhibitors of mitotic spindle in dividing cells have been identified. Vinblastin with these structural changes in connection kinotokor - microtubules and centrosomes in a dividing cell, the mitotic spindle stop (45).Salicylic acid belongs to a group of phenolic compounds found in plants, and today is widely regarded as a hormone-like substance. These classes of compounds act as growth regulators. Humic substances are natural organic compounds that contain 50 to 90% of organic matter, peat, charcoal, rotten food and non-living organic materials are aquatic and terrestrial ecosystems (2).
Materials and Methods: In this experiment, vinca F2 seeds in the mixed 4: 1 perlite and peat moss to the planting trays were sown. The seedlings at the 6-leaf stage were transfered to the main pot (pot height 30 and 25 cm diameter) The pots bed soil mix consisting of 2 parts soil to one part sand and one part peat moss (v / v) were used and after the establishment of seedlings in pots every two weeks with. Salicylic acid and humic acid concentrations 0 (control), 10, 100, 500 and 1000 mg were treated as a foliar spray. Salicylic acid and humic acid used in the Merck has the solutions according to plant size in proper volume has been prepared and will be sprayed on aerial spraying. According to the bootblack periwinkle flowers and leaves to prevent leaf burn and create the solution at one point, for every cc100 solution, two drops of Tween 20 was added to the solution, then spray on the leaves and leaf fire does not spread. The experiment was conducted in a completely randomized design with 10 replicates at the end of the results by the SAS software analysis and comparison of means by Duncan's multiple range tests was performed.
Results and Discussion: According to the results of the data analysis of different treatments significant impact on the level of 1% of the height, number of leaves, chlorophyll, stomatal conductance, pods and stems of the side. Also, the tally was significant at 5%.According to the results of the data analysis of different treatments on stem diameter had no significant effect. Results of comparing the average of the data showed that treatment with 10 and 500 mg/l of salicylic acid per liter respectively in the first and second measurement and control showed lowest height. Treatment of 100 mg/l of humic acid maximum height was measured in two stages. The results of the comparison showed that an average of 500 mg/l of salicylic acid in a two-step measurement and control had the lowest number of leaves. Treatment with 10 mg/l in the first stage of the operation (L1) and treated with 100 mg/l of humic acid in the second vector data (L2) had the highest number of leaves. Treatment with 10 mg/l of salicylic acid and 100 mg/l of humic acid had the highest chlorophyll. The treatment of 10 mg/l of salicylic acid and 100 mg/l of humic acid had the highest stomatal conductance. The results of the comparison average showed that the 500 mg/l of salicylic acid and humic acid had the greatest impact on the number of flowers. As well as 500 mg/l salicylic acid and humic acid had the greatest impact on the number of pods. The results showed that treatment with 1000 mg/l salicylic acid and humic acid had the greatest effect on stem diameter.
Conclusion: The results of this study indicated that low concentrations of salicylic acid increased plant height, the number of leaves, chlorophyll content and stomatal conductance, which can increase plant resistance against unfavorable environmental conditions.
As a result, the plants treated with salicylic acid can be increased two driven in adverse environmental conditions. The treatment of humic acid by increasing the rate of photosynthesis and increases the amount of material available for plant growth. This increase can accelerate the growth of the main branch and side periwinkle plant medicinal plants and enhances the appearance of the plant.
