Medicinal Plants
Yunos Pourbeyrami Hir; Esmaeil Chamani; Mahsa Ahadzadeh; Shabnam Shaker; Roghayyeh Nabipour Sanjbod
Abstract
Introduction
The use of medicinal plants has surged in recent times, with a substantial portion of modern medicines derived from botanical sources. This surge in demand underscores the potential of cultivating and producing medicinal plants to not only bolster public health but also significantly contribute ...
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Introduction
The use of medicinal plants has surged in recent times, with a substantial portion of modern medicines derived from botanical sources. This surge in demand underscores the potential of cultivating and producing medicinal plants to not only bolster public health but also significantly contribute to a society's economic well-being.Salvia nemorosa is an herbaceous, perennial, and medicinal plant. This valuable plant belongs to the Labiatae family. The aerial parts of the plant, especially the leaves, contain essential oils. Furthermore, S. nemorosa contains chemical compositions such as saponin, organic acids, diterpenes, triterpenes, polyphenols, and a bitter substance called picrosalvin. In the plant tissue culture, the preparation of culture medium and formulation compatible with the tissue of the tested plant is vital for efficient regeneration and plant growth. Generally, no specific culture medium can be recommended for the growth of explants, and is necessary to make changes in the culture medium in order to better respond to different types of explants. Carbon nanotubes are nanomaterials that can be added to the culture medium and increase the growth rate of different parts of the plant such as roots, stems, and branches. Cytokines also commonly participate in cell divisions and proliferate shoots by removing terminal dominance. Considering the mentioned importance, this study was conducted to investigate the effect of carbon nanotubes, kinetin, and their interactions on some morphophysiological and phytochemical properties of S. nemorosa under in vitro conditions.
Materials and Methods
This experiment was carried out in the tissue culture and biotechnology laboratory of the Department of Horticultural Sciences, Faculty of Agriculture, University of Mohaghegh Ardabili. This study was conducted based on factorial design (completely randomized design) with five replications. The treatments included five levels of carbon nanotubes (0, 10, 20, 40, and 80 mg/L CNT) and three levels of kinetin (0, 2, and 4 mg/L CK). The seeds of S. nemorosa were sterilized with 70% ethanol for 40 seconds and then 2.5% hypochlorite sodium for 15 minutes (then the seed were washed with deionized water three times 3, 5, and 15 minutes). The sterilized seeds were planted in MS medium containing 30 g/L sucrose and 8 g/L agar, and then transferred to a growth chamber. After 40 days, the obtained seedlings were cut into single nodes by removing leaves and transferred to the main treatment medium. After that, some traits such as fresh weight, number of branches, number of leaves, number of roots, leaf area, root length, seedlings height, viability rate, germination rate, chlorophylls a, b, carotenoid content, phenol content, and flavonoid content were examined. The obtained data related to the experiment were analyzed with SAS software, the comparison of data means was done with Duncan's test, and the graphs were plotted using Excel software.
Results and Discussion
The results indicated that the interaction effect of carbon nanotubes and kinetin on the indices of fresh weight, number of branches, number of leaves, number of roots, leaf area, root length, seedling height, survival percentage, germination percentage, pigment content photosynthesis was not significant. On the contrary, the interaction effect of two treatments on the content of total flavonoid and total phenol showed a significant difference. The highest phenol content was obtained in the treatment combination of 80 mg/L of carbon nanotubes and 4 mg/L of kinetin. On the other hand, increasing the concentration of carbon nanotubes up to 80 mg/L, the average shoot production, number of leaves, leaf area, plant height, root number, root length, chlorophyll a, b, carotenoid, phenol and flavonoid content. increased significantly. Also, by increasing the concentration of kinetin (4 mg/liter), the number of branches, the content of chlorophyll a and b increased significantly.
Conclusion
The characteristics of the S. nemorosa plant, except for fresh weight, significantly increased under the influence of carbon nanotube treatments. However, under the influence of kinetin treatment, only the number of branches, chlorophyll a, b, phenol content, and flavonoid content showed a significant increase. Based on the results of this study, carbon nanotubes can be used for proliferation and increasing the secondary metabolites of S. nemorosa. Despite the results of this study, it is still possible to use higher concentrations of carbon nanotubes in future research in order to increase the phytochemical properties and productivity of other medicinal plants.
Pomology
Hossein Sartip; Ali Akbar Shokouhian; Esmaeil Chamani; Alireza Ghanbari
Abstract
IntroductionSweet cherry is very popular due to its early maturity, high transportability, attractive appearance and good taste of the fruit. The high content of sugars, ascorbic acid, vitamins, carbohydrates and organic acids in the fruit increases the interest in this product both in industrial gardening ...
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IntroductionSweet cherry is very popular due to its early maturity, high transportability, attractive appearance and good taste of the fruit. The high content of sugars, ascorbic acid, vitamins, carbohydrates and organic acids in the fruit increases the interest in this product both in industrial gardening and home gardening. The damage caused by cold in the critical stages of plant growth is one of the important factors in reducing the yield of plants all over the world. Salicylic acid is one of the phenolic compounds that is produced by the roots, and by reducing the activities of reactive oxygen species, it increases the resistance of plants to various environmental stresses (Mahmoudi et al., 2019). Salicylic acid not only plays an important role in determining the quality, color and taste of grape fruit (Hajivand & Rahmati, 2018), but also in the plant's response to environmental stresses such as drought (Miura & Tad, 2014), cold (Kosova et al., 2014) salinity (Noreen et al., 2014) and heavy metal stress (Mahmoudi et al., 2019) are effective. Commercial compounds such as plant growth regulators, including antiperspirant and antifreeze substances, are also used to increase cold resistance or delay the breaking of bud stagnation in horticultural crops (Mahmoudzade et al., 2012). Another way to reduce spring cold damage is to use Natural Plant Antifreeze. These materials either act as a mechanical barrier to prevent the formation of ice crystals on sensitive plant tissues or activate cold resistance systems in the plant (Hajivand & Rahmati, 2018).Materials and MethodsIn order to investigate the effect of the application of growth regulators, on the cold resistance of the cherry tree variety "Siah Daneh Mashhad", a factorial experiment was conducted in the form of a randomized complete block design with 3 factors of growth regulators each at 3 levels. (salicylic acid: zero, 1.5 and 3 mM), (natural antifreeze Thiofer: zero, 2.5 and 5 per 1000) and (soybean oil: zero, 2.5 and 5 per 1000)} and in 4 repetitions It was performed in a commercial garden located in Serain city in 2018 and 2019.Results and DiscussionThe results of the variance analysis revealed that the three-way interactions of the experimental factors significantly influenced the percentage of healthy pistils and the duration of flowering cessation at the 1% probability level. The highest percentage of healthy pistils, reaching 69.25%, was observed in the treatment involving foliar spray application of salicylic acid (1.5 mM) combined with soybean oil (5/1000) and foliar spraying of Thiofer natural antifreeze (5/1000). Furthermore, the combined effect of salicylic acid and natural antifreeze was found to be the most effective in delaying the onset of flower opening. The highest amount of ion leakage percentage was observed in the control treatment and the lowest amount was observed in the 3 mM salicylic acid foliar treatment along with soybean oil (5 per 1000) Thiofer. According to the data variance analysis table (Table 1), the ion leakage index was affected by the simple effect of salicylic acid, soybean oil and antifreeze and the interaction effect of salicylic acid oil × salt, salicylic acid × soybean oil, and soybean oil × antifreeze. The activity of malondialdehyde was affected by the simple effect of salicylic acid and soybean oil and the interaction effect of salicylic acid × year and year × soybean oil (Table 1). The total phenolic content was also affected by the simple effect of salicylic acid and soybean oil (Table 1). According to Figure 11, increasing the concentration of salicylic acid increased the content of total phenol, so that the highest content of total phenol (53.22%) was obtained in the treatment of 3 mM salicylic acid, while there was no significant difference in the treatment of 1.5 mM salicylic acid. . To adapt to the cold, phenolic compounds accumulate in plants, which are related to the antioxidant capacity of the plant (Mozafari &Yazdan Panah, 2018). A decrease in temperature increases the accumulation of phenolic compounds in the plant and can act as a mechanism to adapt and overcome the oxidative stress caused by low temperature (Balasundram et al., 2007). Balasundram and colleagues (Balasundram et al., 2007) noted that grape plants exhibit an accumulation of phenolic compounds and proteins while maintaining membrane stability at low temperatures. This accumulation leads to reduced production of malondialdehyde, enhancing the plant's adaptability and tolerance to cold temperatures, thereby reducing the risk of freezing. Similar observations of increased phenolic compound levels during cold adaptation have been reported in pistachios (Palonen, 1999) and apples (Huang & Wang, 1982). Chen and Tian (Chan & Tian, 2006) reported an increase in phenolic compound accumulation following enhanced activity of phenylalanine ammonia-lyase enzymes in grapes treated with salicylic acid. From their findings, they concluded that salicylic acid plays a pivotal role in the biosynthesis of phenolic compounds and the activation of plant defense genes.ConclusionAccording to the observations of this research, it can be concluded that the use of salicylic acid along with soybean oil and natural antifreeze of Thiofer is a suitable solution in order to delay the opening time of flowers and also to increase the indicators of cherry cold resistance against The tension is cold.
Pomology
Masumeh Jafari; Ali Akbar Shokouhian; Esmaeil Chamani; Akbar Ghavedal
Abstract
Introduction Iron has a significant effect on the quantity and quality of agriculture products. Factors affecting the absorption of this element increase its efficiency. Meanwhile, the pH of the nutrient solution plays an important role in the absorption of iron. Iron is one of the essential elements ...
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Introduction Iron has a significant effect on the quantity and quality of agriculture products. Factors affecting the absorption of this element increase its efficiency. Meanwhile, the pH of the nutrient solution plays an important role in the absorption of iron. Iron is one of the essential elements for plant growth and plays an essential role in chloroplasts. Due to iron deficiency, the activity of several enzymes such as catalase, cytochrome and oxidase and ferroxin is significantly reduced. The amount of iron in the soil is high, but plants only absorb two-capacity of soluble soil, which is negligible compared to the total iron. Soil environmental conditions affect the amount of iron by the plant, so it is difficult to control the uptake of iron by the plant. It has the highest ability to absorb iron and manganese at pH less than 6. For this reason, acetic acid was used to stabilize the pH of the solution. Acetic Acid is a polar solvent and an organic compound. The use of inexpensive organic acid and citric acid in agriculture, despite its positive effects on calcareous soils and their reasonable price is still not common in Iran. Acetic acid has a carboxylic group and therefore has all the properties of an acid.Materials and methods This experiment was carried out in the form of double split-split plot in a completely randomized block design on a strawberry plant of diamant cultivar in the research station of Mohaghegh Ardabili University during the years 2015- 2017. Factors included Acetic Acid (0, 1, 2 and %3), iron in two levels (Sequestrin 25gr and Nano 1gr) along with two levels of agricultural soap agents (0 and % 7/5) as foliar spraying. Foliar application of pH nutrient solution from the three-leaf stage of plant (mid-April) to the end of May a total of five times 10 days apart in both years. Two weeks after the last foliar application (June of the second year) plant growth indices including, total chlorophyll, number of fruits per plant, fruit pH, total acidity, soluble solids, fruit volume, fruit fresh weight, fruit dry weight, and leaf iron content and fruit yield per plant was measured.Result The results of this study showed that the three-way interaction of foliar application of Acetic Acid with iron with agricultural Soap on the average concentration of leaf iron and number of fruits at the probability level was %5 and in total chlorophyll at the %1 probability level was significant, the best result in the mentioned traits was the combination of treatment of Acetic Acid %2 with Sequestrin iron with soap Moyan (%7/5). The interaction effect of Acetic Acid and iron type on traits of fruit pH, fruit volume, fruit fresh and dry weight at %5 probability level and in yield at %1 probability level were significant. Fruit volume, fruit fresh weight and fruit dry weight and yield showed the best results due to treatment combination of %2 Acetic Acid with Sequestrin iron fertilizer. But for fruit pH trait, the best results were related to the treatment of Acetic Acid %3 on the leave of Sequestrin iron. The Interaction of the iron and agricultural Soap was significant on the amount of soluble solids at %5 probability level and the total acidity strawberries fruit was significant at %1 probability level. In the case of soluble soild and Acidity, Sequestrin iron treatment and agricultural soap had the best results. According to the results of this study, the combination %2 Acetic Acid with of iron fertilizer Sequesterin in combination with agricultural soap Moyan (%7/5) due to the reduction in leaf area tensions improves the obsorption of iron and the quantitative and quality characteristics strawberries.Conclusion Based on the findings of this research, it can be concluded that Acetic acid %2, has better result on the absorption of iron fertilizer, along with agricultural soap (% 7/5) application than other treatments and qualitative traits of strawberry fruit. In treatment %2 Acetic acid most measured traits such as total chlorophyll, leaf iron, fresh weight, fruit dry weight and yield has shown a significant increase. Acetic acid by reducing the pH of the cell sauce and increasing the activity of the reductase enzyme increase the iron solubility and easily provides the iron to the plant. Due to the pH alkalinity of most soils in Iran, the use of Acetic acid in iron nutrient solution on the absorption of iron Sequestrin in combination with % 7/5 of agricultural soap due to having a hydrophilic head and a hydrophobic with %40 to %50 increase in effective of foliar fertilizers. As a result, with more absorption it can be an advanced approach to better absorption of iron by plant and increase the quantity and quality in the product.
Roghayeh Fathi; Mehdi Mohebodini; Esmaeil Chamani
Abstract
Introduction: Transformed hairy roots obtained by infection of plants with Agrobacterium rhizogenes strains are used as a tissue culture tool for secondary metabolite production since hairy roots are genetically and biologically stable and they are able to produce metabolite within a short time. Chicory ...
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Introduction: Transformed hairy roots obtained by infection of plants with Agrobacterium rhizogenes strains are used as a tissue culture tool for secondary metabolite production since hairy roots are genetically and biologically stable and they are able to produce metabolite within a short time. Chicory (Cichorium intybus L.) is a medicinal plant from Asteraceae. This plant contains many important metabolites include chicoric asid, inulin, scoline, coumarin and flavonoids.
Materials and Methods: The seeds were surface-sterilized by rinsing with 2% benomil for 30 min and later sterilized with 5% sodium hypochlorite for 20 min and subsequently with 70% ethanol for 90 s. the wounded leaf of 20 and 28-day-old seedlings inoculated with Agrobacterium rhizogenes 11325. Bacterial colonies were cultured on liquid LB medium for 1 day. The explants were submerged in the solution for 15 min and were shaken gently. After that, the explants were drained on sterile filter paper and then transferred to MS solid medium. The explants were incubated at (25 ± 2) ◦C under dark condition for three different co-cultivation period (24, 48 and 72 hours). In the next step, the explants were transferred to MS solid medium supplemented with 500 mg/L cefotaxime. The explants were incubated at (25 ± 2) ◦C under a 16-h photoperiod condition for four weeks. At the end of incubation time, the percentage of hairy root induction, root number and root length were investigated. Genomic DNA of Cichorium intybus L. hairy root was extracted following the method of CTAB and subjected to PCR analysis. Non-transformed root DNA was used as a negative control and pRi11325 plasmid DNA was used as positive control. The pair of primers specific to the rolB fragment sequences was: 5-ATGGATCCCAAATTGCTATTCCCCACGA -3 and 5-TAGGCTTCTTTCATTCGGTTTACTGCAGC-3. The amplification protocol for the rolB was a 5 min melting at 94 ◦C followed by 35 cycles of a 5 min melting at 94 ◦C, a 45 s annealing at 55 ◦C and a 1 min elongation at 72 ◦C, and final elongation at 72 ◦C for 7 min. PCR products were electrophoretically separated on 0.8 % agarose gels (w/v) and stained with gel red. To determine the best media composition for growth of hairy roots, approximately 100 mg FW roots were cultured in basal liquid MS, solid MS and liquid 1/2MS medium supplemented with 500 mg/L cefotaxime on a rotary shaker (90 rpm) at 25 ◦C in dark for 5 weeks, and increase in fresh weight and dry weight was recorded at 5 week later. Three replicates were made for each experimental set. Non-transformed roots were cultured on same mediums.
Results and Discussion: Each of the two different explants – leaf and petiole showed different levels of hairy root induction in different co-culture time. Hairy root induction was observed 7 days after co-cultivation. There were low adventitious roots formed from control explants. 72 hours co-cultivation period was found to be more efficient in inducing hairy root phenotype in both explant types. The infection of explants by A. rhizogenes is a process that implies a succession of events, including the transfer and integration of T-DNA, into the host plant genome. Therefore, the time of contact between A. rhizogenes and the explants determines the transformation efficiency. A maximum of transformation frequency (53.33%) was observed in leaf explants in 72 hours co-cultivation followed by 33.33% in petiole explants in 72 hours co-cultivation. In leaf explants maximum root number (8.5 roots per explant) and maximum length of root (9.16 cm) induced from 20-day-old seedling in 72 hours co-culture. In petiole explants maximum root number (3.66 roots per explant) and maximum root length (11.46 cm) induced from 28-day-old seedling in 72 hours co-culture. The “hairy root” phenotype characterized by high branching and fast growth was previously reported in “reactive species” like Datura innoxia but not for “difficult species”. The high phenolic content of woody plant species could be responsible for the reduced hairy root phenotype as reported for gingko or pine. Three different culture media (solid MS, liquid MS and 1/2 MS media) were tested to determine the best suitable media for the maximum Biomass of hairy roots line.
Conclusions: Genetically transformed hairy roots obtained by infection of plants with Agrobacterium rhizogenes are suitable source for production of bioactive molecules due to their genetic stability and generally show fast growth in culture media free of growth hormones. This study describes the protocol for hairy root induction which could further be useful for the production of secondary metabolites and biomass.
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
Mehdi Mohebodini; Zahra Azimzadeh; Esmaeil Chamani; Malihe Erfani
Abstract
Introduction: Lily (L. ledebourii) is the rarestspeciesof thegenusLilium, and grows in Caucasus region. Iranis one of the important distribution areas of this endangered species. It is important as an ornamental plant due to its large and attractive white flowers that are equal to those of commercial ...
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Introduction: Lily (L. ledebourii) is the rarestspeciesof thegenusLilium, and grows in Caucasus region. Iranis one of the important distribution areas of this endangered species. It is important as an ornamental plant due to its large and attractive white flowers that are equal to those of commercial lilies in terms of beauty.The two main constraints on growing this plant are a low multiplication rate and the high cost of bulb production. Five to ten flowers commonly appear on each plant, even specimens with up to 15 flowers have been observed. Plant tissue culture techniques are widely used in plant propagation and using these methods can effectively provide micro-propagation of this plant in large scale. High percentage ofregeneration is necessary for plant protection, using in the breeding programs and gene transfer to this plant. Therefore, the effect of plant growth regulators and abiotic stress (ultrasound) werestudied on the bulblet production and root induction of Lilium ledebourii.
Materials and Methods: The experiment was factorial based on completely randomized design with four replicattions and was carried out in tissue culture lab of University of MohagheghArdabili in 2015. For this purpose, segmentsof scale explant was treated with ultrasound and cultured on MS medium supplemented with different concentrations of NAA and BA alone and/or in combination with each other. In this experiment, different concentrations of NAA (0, 0.01, 0.1 and 1 mgl-1) and BA (0, 0.01, 0.1 and 1 mgl-1) and different Ultrasound exposure duration (0, 5, 10, 20 and 30 second) were studied. In order to remove possible contamination from the media, all media were autoclaved for 20 minutes at 121 °C. At the end of the experiment, the number of bulblet, root length, fresh weight of bulblet were recorded. The cultures were kept at 25±2°C under illumination with daylight fluorescent lamps (30 mol m-2s-1) at 16 h photoperiod. Data was subjected for analysis of variance and compare means using SPSS 16.
Results and Discussion: The results showed that ultrasound had negative effect onroot length, so that the highest root length was observed in explants without ultrasound treatment. Result also indicated that ultrasound had positive effect on bulblet production and root induction. A different effect of growth regulators was observed in similar media on the bulblet formation percentage. The 0.1 NAA concentration had a higher efficiency while increasing NAA insignificantly decreased bulblet induction. The highest total weight and number of bulblets obtained by 0.1 mgl-1 NAA. Concentrations of NAA increased rooting percentage. Different concentrations of NAA had also significant effects on some traits. So that, the highest weight of bulblets obtained by 0.01 and 0.1 mgl-1 BA and the highest number of roots obtained in control. Bulblet maximum mean weightwas in30 seconds ofultrasoundtreatment, which hada significantdifference with the control treatment (without ultrasoundtreatment). In the other hand, ultrasound increased the number and weight of bulblets.Mechanical stress and microstreaming by acoustic cavitation might be considered as the most possible cause of the various physiological effects of ultrasound on cells. The enhancement of V-ATPase transport and ATP hydrolysis activities seem to be an ultrasound-induced metabolic response of cells. High-intensity ultrasound is well known to be destructive to biological materials, disrupting the cell membranes and deactivating biological molecules such as enzymes and DNA. Low-intensity ultrasound, on the other hand, has shown a range of sub lethal biological effects that are of potential significance in biotechnology. There are several processes that take place in the presence of cells or enzymes activated by ultrasonic waves. High-energy ultrasonic waves break the cells and denature the enzymes. Low-energy ultrasound can modify cellular metabolisms or facilitate the uptake of nutrient, and make them easily through the cellular walls and membranes. In the case of enzymes, the increase in the mass transfer rate of the reagents to the active site seems to be a most important factor.
Conclusions: The results showedthatthebulblet production at first stages and a little root formation in tissue culture is useful for fast bulblet inductionandthenrooting. Finally, it seems that ultrasound in combination with plant growth regulators have the potential to produce the highest average number of bulblets in the scale explant.
Esmaeil Chamani; Marziyeh Ghamari; Mahdi Mohoboldini; Alireza Ghanbari; Hamid Reza 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
Hassan Hasani Jifroudi; Mehdi Mohebodini; Behrooz Esmaielpour; Esmaeil Chamani
Abstract
Introduction: Fenugreek (Trigonella foenum- graecum) is a medicinal plant extensively distributed in most regions of the world. Fenugreek is an annual plant from the family of papilionaceae, leguminosae. Fenugreek leaves and seeds have been used extensively to prepare extracts and powders for medicinal ...
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Introduction: Fenugreek (Trigonella foenum- graecum) is a medicinal plant extensively distributed in most regions of the world. Fenugreek is an annual plant from the family of papilionaceae, leguminosae. Fenugreek leaves and seeds have been used extensively to prepare extracts and powders for medicinal uses. Its root, leaf and seed contain a number of important medicinal compounds such as polysaccharide, galactomannan, different saponins such as diosgenin, yamogenin, mucilage, volatile oil and alkaloids such as choline and trigonelline. Plant tissue culture is fundamental to most aspects of biotechnology of plants. Establishment of an efficient callus induction and direct regeneration protocol is an essential prerequisite in harnessing the advantage of cell and tissue culture for genetic improvement. For the successful application of the tissue culture technique in plant breeding, callus induction and plant regeneration potential of each plant must be determined. The present study was performed in order to determine the optimum concentration of plant growth regulators (IBA + TDZ) for producing of in vitro plantlet using cotyledon and hypocotyl as an explant for two different Iranian genotypes (Ardestani and Neyshabouri).
Materials and Methods: In this investigation, Ardestani and Neyshabouri genotypes were used for callus induction and direct shoot regeneration. The medium used in this investigation was MS (Murashige and Skoog) basal medium. Then seeds were germinated on MS medium. For callus induction and direct shoot regeneration, cotyledon and hypocotyl explants were excised from 8-day-old sterile seedlings and cultured on MS medium containing various concentrations of IBA and TDZ. In this experiment, two combinations (TDZ + IBA) were used. In the first composition, IBA had four levels (0, 0.1, 0.3, 0.5 mg l-1) and TDZ had five levels (0, 0.2, 0.4, 0.6, 0.8 mg l-1) and in the second composition, IBA had four levels (0, 0.05, 0.1, 0.15 mg l-1) and TDZ had seven levels (0, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45 mg l-1).The experimental designs were factorial based on completely randomized design with four replications. Cultures were incubated at 25° C ± 2 with a 16/8 hour (day/night) photoperiod and an irradiance of 1500 LUX using Sylvania cool white fluorescent tubes. The percentage of callus induction, direct shoot regeneration and average weight of callus were calculated for cotyledon and hypocotyl explants. All Data were analyzed using SPSS16, and mean comparisons were performed with duncan’s multiple range test (P < 0.05).
Results and Discussion: According to our results, explants cultured on MS basal medium without plant growth regulators (control) produced no callus. However, after two weeks, callus formed in both of Ardestani and Neyshabouri genotypes from cotyledon and hypocotyl explants in the presence of IBA + TDZ plant growth regulators in most of the combinations. In hypocotyl explants of Neyshabouri genotype, the highest callus induction was obtained from the medium containing 0.15 mg l-1 IBA + 0.45 mg l-1 TDZ (96.87%). Various important factors such as genotype, source of explants and plant growth regulators significantly influence direct regeneration. Direct regeneration was obtained from hypocotyl explants for Neyshabouri genotype in combination IBA + TDZ. The highest percentage of direct shoot regeneration was observed in MS medium containing 0.05 mg l-1 IBA + 0.35 mg l-1 TDZ in hypocotyl explants of Neyshabouri genotype (37.5%). Direct shoot regeneration requires plant cells to undergo dedifferentiation which is known to be affected by not only exogenous plant growth regulators but also endogenous content of the hormones. Different tissues may have different levels of endogenous hormones and, therefore, the type of explant source would have a critical impact on the regeneration success. In our study, when cotyledon and hypocotyl explants were compared, it was clear that hypocotyl explants were much more productive for direct shoot regeneration than cotyledon explants.
Conclusions: Callus induction and direct shoot regeneration are as in vitro tissue culture methods. Plant growth regulators and types of explant and genotype are the most important factors for callus induction and direct shoot regeneration phases. Therefore, optimization of these factors is essential to establish a high frequency of callus induction, direct shoot regeneration and gene transferring to this plant. According to the results of this investigation, it is recommended to apply plant growth regulators that were used in this study for other landraces of fenugreek cultured in Iran and select the best genotypes in response to tissue culture conditions for using in future studies.
Parisa Daryani; Naser Zare; Esmaeil Chamani; Parisa Sheikhzade Mosaddegh; Davoud Javadi Mojaddad
Abstract
In this research, the effects of different basal medium and plant growth regulators on in vitro establishment and growth of hazelnut were investigated. For this, the spring apical and auxiliary buds of cv. Fertile were sterilized and cultured on NRM, MS and 1/2MS basal media containing 0.01 mg/l IBA ...
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In this research, the effects of different basal medium and plant growth regulators on in vitro establishment and growth of hazelnut were investigated. For this, the spring apical and auxiliary buds of cv. Fertile were sterilized and cultured on NRM, MS and 1/2MS basal media containing 0.01 mg/l IBA and different levels of BAP. The results indicated that percentage of explant growth (shooting), number of leaves per explant and shoot length influenced significantly by basal media and concentration of plant growth regulator. Based on orthogonal contrasts analysis, although the highest percentage of shooting was obtained on MS medium, shoot length of explants cultured on NRM basal medium were significantly higher than those of MS and 1/2MS. The best growth response of explants in establishment stage (50% shooting, 5.33 leaves per shoot and 1.6 cm shoot length) were obtained with NRM medium supplemented with 0.01 mg/l IBA and 1 mg/l BAP. Shoots derived from establishment stage were cut to single-node explants and transferred on NRM medium supplemented with 0.05 mg/l IBA and different levels of BAP and TDZ. The highest percentage of explants growth with lowest callgenesis and explnt browning were obtained on NRM medium containing 0.05 mg/l IBA and 5 mg/l BAP.
Homeyra Hadizadeh; Mehdi Mohebodini; Behrooz Esmaeilpour; Esmaeil Chamani
Abstract
Introduction: Chicory (Cichorium intybus L.) belongs to Asteraceae family is commonly known as witloof chicory. The leaves and the roots of this medicinal plant are edible and commonly used as salad. Some varieties are also cultivated as coffee substitute after roasting the roots. All parts of the plant ...
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Introduction: Chicory (Cichorium intybus L.) belongs to Asteraceae family is commonly known as witloof chicory. The leaves and the roots of this medicinal plant are edible and commonly used as salad. Some varieties are also cultivated as coffee substitute after roasting the roots. All parts of the plant contain these volatile oils, with the majority of the toxic components concentrated in the plant's root. In folk medicine, the plant is used for the treatment of diarrhea, spleen enlargement, fever, and vomiting. Antihepatotoxic activity on damaged rat’s liver sections and anti-bacterial activity of this crop has been recently reported. In vitro regeneration from leaf explants with various hormonal combinations has been reported previously. Moreover, in vitro regeneration of Chicory from cotyledon explants using different combinations of plant growth regulators has been studied. Also, a protocol for the regeneration of plantlets from leaf and petiole explants of witloof chicory has been developed. The aim of the present investigation was optimization of callus induction and shoot regeneration from leaf and petiole tissues of Chicory (Esfahan genotype).
Materials and Methods: In this investigation, Esfahan genotype was used for callus induction and direct shoot regeneration. Seeds were first washed with running tap water for 30 min then seeds were surface sterilized by dipping in 70% ethanol for 90 s and rinsed with sterile distilled water, followed by immersing in 5% sodium hypochlorite solution for 25 min and thereafter rinsed for 30 min with sterile distilled water. The basal medium used in this investigation was MS. For shoot regeneration, leaf and petiole explants (5 mm segments) were excised from 4-week-old sterile seedlings and cultured on MS medium containing different combinations of NAA / BA and KIN / BA in two separate experiments. Experiments were performed factorial based on completely randomized design. Cultures were incubated at 25° C ± 2 with a 16/8 hour (day/night) photoperiod and an irradiance of 1500 LUX using Sylvania cool white fluorescent tubes. The percentage of callus induction, shoot regeneration and the number of regenerated shoots were calculated for the leaf and petiole explants. Data was subjected for analysis of variance and means were compared in 5% level with Duncan’s multiple range tests.
Results and Discussion: Explants cultured on medium containing either no plant growth regulators (control) or cytokines alone produced no callus. However, after 2 weeks, other concentrations of NAA and BA indicated callus formation from leaf and petiole explants in all hormone combinations. In leaf explants, the highest callus induction were obtained in the medium containing 0.3 mg l-1 NAA with 1 mg l-1 KIN and 0.3 mg l-1 NAA with 1.5 mg l-1 KIN (81.25%). Leaf and petiole explants cultured on medium containing no plant growth regulators (control treatment) and medium containing NAA produced no shoots. The combination of 0.3 mg l-1 NAA and 0.1 mg l-1 BA was the best treatment tested. This treatment produced 2.7 shoots per explant at 71% shoot regeneration frequency in leaf explant and 2.73 shoots per explant at 73% shoot regeneration frequency in petiole explants. The results also showed that the highest percentage of regeneration and the highest number of regenerated shoots were obtained in the medium containing 0.1 mg l-1 NAA and 1 mg l-1 KIN in leaf explants (65.6% regeneration and 1.37 shoots per explant, respectively). The highest number of regenerated shoots was obtained in the medium containing 0.3 mg l-1 NAA and 0.5 mg l-1 KIN in petiole explants (40.6% regeneration and 0.5 shoot per explants, respectively. Shoot regeneration requires plant cells to undergo dedifferentiation which is known to be affected by not only exogenous plant growth regulators but also endogenous content of the hormones. Different tissues may have different levels of endogenous hormones and, therefore, the type of explant source would have a critical impact on the regeneration success. In our study, when leaf and petiole explants were compared, it was clear that leaf explants were much more productive for regeneration than petiole explants.
Conclusion: Callus induction and shoot regeneration are in vitro tissue culture methods. Plant growth regulators and types of explant are the most important factors for callus induction and shoot regeneration phases. Therefore, optimization of these factors is essential to establish a high frequency of callus induction, shoot regeneration and gene transfer to this plant.
Esmaeil Chamani; Mohammad Bonyadi; Alireza 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.
Esmaeil Chamani; Mobina Taheri
Abstract
Three experiments were conducted in tissue culture and biotechnology laboratory of Horticulture Department of Mohaghegh Ardabili University in 2012. For the regeneration of plant from seed, different concentrations of NaOH (5, 10, 15, 20 M) and various scarification methods with sandpaper (soft scarification, ...
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Three experiments were conducted in tissue culture and biotechnology laboratory of Horticulture Department of Mohaghegh Ardabili University in 2012. For the regeneration of plant from seed, different concentrations of NaOH (5, 10, 15, 20 M) and various scarification methods with sandpaper (soft scarification, medium scarification and hard scarification) were used based on completely randomized design with 4 replications. The results of experiment revealed that seeds treated by 20 M NaOH and hard scarification produced the highest germination rate. After 2 months of seed germination, hypocotyles of seeds were used as explants and cultured in MS medium containing different concentration of 2,4-D, picloram, TDZ and BA (1, 2, 4 mg/l) based on completely randomized design with 4 replications. Mean comparison revealed that explants treated by 4 mg/l picloram and 1 mg/l 2,4-D produced the highest callus content. Mean comparison showed that explant treated by 1 mg/l BA produced the highest shoots. However, to investigate the soluble protein changes during growth stages and to study the effects of 2,4-D, picloram, TDZ and BA on soluble protein and experiment was conducted. The result showed that by increasing the plant age, soluble protein was reduced and also the highest soluble protein was found after 4 weeks of germination. The result also showed that explants treated by 4 mg/l picloram and 1 mg/l BA produced the highest soluble protein content.
Esmaeil Chamani; Behrooz Esmaeilpour; Yunos Pourbeyrami Hir; Hassan Maleki Lajayer; Akbar Saadati
Abstract
Two separated experiments were conducted to evaluate the effects of different concentrations of thidiazouron (10, 20, 30, 40 and 50 µM) and humic acid (1, 10, 100, 1000 and 10000 ppm) on vase life of cut Alstroemria flowers. Experiments were carried out based on completely randomized design with 8 replications ...
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Two separated experiments were conducted to evaluate the effects of different concentrations of thidiazouron (10, 20, 30, 40 and 50 µM) and humic acid (1, 10, 100, 1000 and 10000 ppm) on vase life of cut Alstroemria flowers. Experiments were carried out based on completely randomized design with 8 replications in postharvest laboratory of Horticultural Department, Mohaghegh Ardabili University in 2009. The results showed that lower concentrations of humic acid had positive effects on flower vase life, water content of tissue and solution uptake, while didn’t affect relative fresh mass and leaf chlorophyll content. In both experiments results also indicated that humic acid in higher concentrations affected all traits, negatively. Howewer, this compound at 10000 ppm decreased flower vase life, water content, relative fresh mass, solution uptake and chlorophyll content. By increasing the humic acid concentration up to 100 ppm the vase life of cut flowers improved. Compared with control and other treatments, 1000 ppm of humic acid decreased flower vase life, considerably. The highest vase life of flowers devoted to 10 µm of TDZ, however higher concentrations of this compound reduced flowers vase life. Moreover, the highest solution uptake and leaf chlorophyll content obtained by 30 µm TDZ, while the highest relative fresh mass devoted to 40 µm of this compound.
