ahmad sharifi; fatemeh keykha; mahboobeh yazdi; abdolreza bagheri
Abstract
Introduction: Lily (Lilium spp.) is a genus of herbaceous flowering plants, which consisted of many beautiful ornamental species with large prominent flowers. Most species are native to the Northern hemisphere temperate, though their range extends into the Northern subtropics. Some specific hybrids of ...
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Introduction: Lily (Lilium spp.) is a genus of herbaceous flowering plants, which consisted of many beautiful ornamental species with large prominent flowers. Most species are native to the Northern hemisphere temperate, though their range extends into the Northern subtropics. Some specific hybrids of Lilium spp. have been developed as cut flower in controlled conditions and in some cases can be grown as pot plant. Propagation rate of lily in natural clonal propagation methods is very low and one year produces of 1-2 bulblets per bulb scale. There is also possibility of disease transmission; so that, tissue culture techniques has provided an efficient method for its micropropagation.
Materials and Methods: In this study, two separate experiments under In vitro conditions the bulblet regeneration from thin cell layer (TCL) explants of Lilium spp. was investigated. In the first experiment, after two months the effect of TCL explants with 1, 3 and 5 mm thickness on MS medium containing 1 mg/l BA, 2ip and kin in combination with 0.5 mg/l NAA on regeneration parameters were assayed. In the second experiment to determine the effect of cultivar and cytokinin types, 3mm thickness TCL explants of five cultivars (Robina, Donato, Nymph, Lessoto and Roxana) were tested on MS medium containing different plant growth regulator (PGR) compounds including BA, kin, 2ip and TDZ at concentration of 1 mg/l in combination with 0.5 mg/l NAA. The regeneration parameters were assayed after four months. In all experiments, the medium was adjusted to pH 5.8 and autoclaved at 121°C for 15 min. All cultures were incubated at 25 ± 2°C with a 16 h photoperiod under cool white flourescent lights (30 µmol/m2).
Results and Discussion: According to the first experiment results, plant growth regulator of BA in all of surveyed parameters except root number was better than other PGRs and explants with 3 mm thickness was the best in all of parameters. The interaction of PGR and explants was significant, however maximum bulblet regeneration was observed in TCL explants with 3 mm thickness in all of PGR treatments (100%). While 1 mm thickness TCL in 2ip and 1 and 5 mm thickness TCL in Kin had the least regeneration percentage. Results revealed that the interaction of explants and medium is a key factor for suitable establishment, regeneration and growth of TCLs. Bulb dormancy is one of the limiting factors in regeneration of bulbous crop species. It seems under In vitro condition explants size and PGR combination of media especially cytokinin affected on breaking of dormancy. Maximum number of leaves and dry weight of bulblets in medium containing BA was significantly higher compared with other treatments. Most of studies confirmed the positive effect of BA on regeneration of lily. The function of cytokinin in plant promoted cell division and differentiation, which lead to growth and maintaining cells in meristematic status.
Result of second experiment showed that cultivar was one of the effective factors on regeneration trait. Oriental lily cultivar "Roxana" had the highest number of roots, bulblets, dry weight and length of plantlets and "Nymph" cultivar showed the lowest percentage of regeneration, dry weight, length of plantlets and rooting obtained. In all of cultivars BA induced more organogenesis percentage and plantlet dry weight, while TDZ induced more rooting percentage.The interaction of cultivar and PGR treatments on percentage of regenerated bulblets and rooting were significant. "Nymph" cultivar had minimum percentage of regeneration and rooting in medium containing TDZ and Kin. Furthermore, "Roxana" cultivar in medium containing BA showed the best dry weight comparison to other treatments.
Conclusion Lily has widely used in the floral industry as a cut flower or potted plant. In recent years, tissue culture was developed as reliable and highly effective method to overcome its limitations of vegetative propagation. The most advantage of this method is high multiplication rate and disease free propagation. In this study, bulblet regeneration of lilium Spp. from TCL explants under in vitro condition was considered as a highly efficient procedure for its micropropagation. With optimization of TCL system some parameters such as exogenously applied plant growth regulators, cultivar, explants types were investigated. Favorable conditions for bulblet regeneration were achieved with 3 mm thickness TCLs in MS medium containing 1 mg/l BA with 0.5 mg/l NAA. This protocol can be used for rapid micropropagation of many cultivars.
Nasrin Farhadi; Saeideh Alizadeh Salteh
Abstract
Introduction: Allium hirtifolium commonly known as Persian shallot is an important wild medicinal plant from Alliaceae family. Persian shallot commonly known as mooseer in Iran is a perennial diploid plant that is native to Iran and grows as a wild plant throughout in the Zagross Mountains range, western ...
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Introduction: Allium hirtifolium commonly known as Persian shallot is an important wild medicinal plant from Alliaceae family. Persian shallot commonly known as mooseer in Iran is a perennial diploid plant that is native to Iran and grows as a wild plant throughout in the Zagross Mountains range, western and southwestern Iran. It is a bulbous herb and usually consists of a single main bulb or rarely two bulbs. Each bulb has a weight of about 8-15 times of a garlic clove. The bulbs of mooseer has been widely used as a traditional herb and spice plant, added to a variety of foods such as salads, pickles, yogurt and different sauces. Conventionally, Persian shallot propagates through bulbs and seeds but these two methods are not commercially efficient due to low growth rate of bulbs and deep dormancy, low viability and germination rate of seeds. In addition, the natural habitat of this plant is under increasing pressure as a result of excessive incorrect harvest that caused to damage the plant density in Iran rangelands. So, improving the efficiency of A. hirtifolium propagation is necessary. A number of positive effects on the growth and productivity of some plants through cytokinin application have been registered by earlier research. The current study aimed to evaluate the effects of pretreatment and foliar application of forchlorfenuron as a safe cytokinin on improving the bulb production, phytochemical compounds and antioxidant attributes of Persian shallot.
Materials and Methods: This experiment was done at research green house of Tabriz University in 2015-2016. For pretreated of Persian shallot bulbs, they were soaked in 0, 50 and 10 mg l-1 forchlorfenuron solutions for 24 h. Then they were cultured in pots contained perlite and vermicompost with 3:1 ratio. Foliar application was applied 2, 4 and 6 weeks after culture with 0, 50, 100 and 150 mg l-1 concentrations of forchlorfenuron. At the end of growth season the number of leaves, number of bublets, fresh and dry weight of bulblet were recorded. The phytochemical compound (protein, phenol and allicin), antioxidant enzymes (catalase, peroxidase, ascorbate peroxidases and superoxide dismutase) and antioxidant activity of bulbs were assayed with spectrophotometry methods.
Results and Discussion: Foliar applications of plant growth regulators such as cytokinins in agriculture crops are reported to be useful in controlling multiple physiological processes, including flower initiation, shoot elongation, bulb production, fruit set and as well as affected the quality characters of products. In this study despite the bulblets number that did not influence by treatments, pretreatment and foliar application of forchlorfenuron significantly increased the leaves number, fresh and dry weight of bulbs in comparison with control plants. The highest leaves number (4.49 per plant) was obtained from pretreatments. The highest fresh weight (91.77 g) was recorded at 5 and 10 mg l-1 pretreatment and 100 mg l-1 (91.63 g) foliar application. The interaction effect of treatments on dry weight was significant and the highest dry weight (19.75 g) was recorded at 10 mg l-1 pretreatment with 100 and 150 mg l-1 foliar application. Allicin content did not show significant variation between treatments and in average was 0.859 mg g-1 FW. Total phenol content significantly influenced by treatments and the highest phenol content (1.585 mg GAE g-1 FW) was recorded at 5 mg l-1 pretreatment with 100 mg l-1 foliar application. The antioxidant enzymes included catalase, peroxidase and ascorbate peroxidase that showed significant increasing under forchlorfenuron treatments. Due to significant effects of forchlorfenuron on antioxidant compounds and enzymes of Persian shallot bulbs, the assay of antioxidant activity also showed a significant increasing in treated bulbs. The maximum percent of antioxidant activity (74.522) was obtained from 100 mg l-1 foliar application. Exogenous application of cytokinins plays an effective role by protecting the fluidity and integrity of plant cell membranes. They properly mediate enzymatic (SOD, APX, and CAT) and non-enzymatic machinery with the result of preventing cell membrane damage by oxidative stress.
Conclusions: Considerable improvement in biochemical and antioxidant attributes of Persian shallot was recorded with pretreatment and foliar application of forchlorfenuron. The present data support the potential uses of the forchlorfenuron for improving the production of weighty bulbs with the high antioxidants attributes in Allium hirtifolium. Pretreated and foliar application at 5 mg l-1 and 100 mg l-1 concentrations of forchlorfenuron, respectively showed the best results and is recommendable for A. hirtifolim production.
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
