Biotechnology
Seyedeh Mohaddeseh Hosseini; Seyed Ali Andi; Amir Sahraroo; Mohammad Bagher Farhangi
Abstract
Background and objectives: Adventitious root cultures of medicinal plants represent a bountiful reservoir of valuable secondary metabolites. The esteemed medicinal plant Echinacea purpurea (L.) Monech is highly sought after for its phytochemical properties, with a traditional ...
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Background and objectives: Adventitious root cultures of medicinal plants represent a bountiful reservoir of valuable secondary metabolites. The esteemed medicinal plant Echinacea purpurea (L.) Monech is highly sought after for its phytochemical properties, with a traditional use of its roots in herbal medicines and dietary supplements as an immune system stimulant for treating inflammatory, viral, and respiratory diseases. However, the heterogeneous synthesis of bioactive compounds in natural environments and the limitations of conventional plant production methods necessitate exploring alternative approaches for obtaining active plant substances and organs. Among various in vitro cultivation techniques, the cultivation of differentiated tissues, specifically roots, consistently demonstrates the ability to yield higher quantities of metabolites than undifferentiated tissues such as callus and suspension cells. Methodology: The primary aim of this study was to accurately determine the ideal combination of plant explants and plant growth regulators that would facilitate the successful establishment of adventitious root suspension cultures. Three distinct plant explants, namely leaf, petiole, and root, were carefully evaluated in this study. These explants were subjected to varying concentrations (0, 0.5, 1, 1.5, and 2 mg.L-1) of two types of plant growth regulators: indole-3-butyric acid (IBA) and 1-naphthalene acetic acid (NAA). After six weeks, the number of adventitious roots developed by the explants under different concentrations of plant growth regulators was measured. The impact of different sucrose concentrations (10, 20, 30, 40, 50, and 60 grams per liter) in the culture medium was investigated over a four-week cultivation period. Various dynamic characteristics of root growth, including fresh weight, dry weight, volume of the remaining medium, electrical conductivity, and pH, were closely examined. Furthermore, phytochemical properties such as total phenol, total flavonoid, and antioxidant activity were assessed. To administer the sucrose treatments, 0.35 grams of adventitious roots were placed inside 250 ml Erlenmeyer flasks containing 50 ml of half MS culture medium, 1 mg.L-1IBA, and varying concentrations of sucrose, maintaining a pH of 5.8. Measurements were taken from three flasks of each treatment weekly. The experiments followed a Completely Randomized Factorial Design, with statistical analysis conducted using SPSS software.Results: Leaf explants exhibited superior root formation to root and petiole explants. The average number of adventitious roots in leaf explants reached 3.37, while petiole and root explants produced only 0.23 and no adventitious roots, respectively. The most effective root formation occurred in MS medium with 1 mg.L-1 IBA, resulting in an average of 4 roots per explant. Hormone concentrations of 0 and 2 mg.L-1 did not induce root formation. In the fourth week of cultivation, the highest fresh weight recorded was 29.8 grams per liter, marking a remarkable 3.4-fold increase from the initial inoculation weight. The culture medium's electrical conductivity and pH decreased as the cultivation progressed. Cultivation weeks did not significantly impact total flavonoid production and related antioxidant activity in adventitious roots. However, the cultivation period significantly affected the total phenol content at 57.56 mg of gallic acid per gram of dry weight. Altogether, lower sucrose concentrations proved more efficient for biomass production, bioactive compound content, and antioxidant activity. The highest biomass accumulation and total phenol content occurred at 3% sucrose. In comparison, in the fourth week of cultivation, the highest total flavonoid content and the highest free radical inhibition percentage were observed at 2% and 1% sucrose, respectively. Notably, a negative correlation was observed between the electrical conductivity of the remaining culture medium and root biomass in lower sucrose concentrations as the culture duration increased.Conclusion: In light of the immense potential harbored by adventitious root suspension cultures of E. purpurea as an alternative source for the production of secondary metabolites, particularly caffeic acid derivatives, the discoveries made in this research have the potential to advance the field of mass production for active root organs, thereby facilitating the synthesis of these precious compounds through the implementation of appropriate laboratory bioreactor systems.
Improvement and breeding
M. Sarafraz; H. Dehghanzadeh; A.F. Hashemi
Abstract
To investigate the effects of biological and chemical phosphorus fertilizers, and gibberellic acid (GA3) on yield and its components, and essential oil of Salvia officinalis L., a factorial experiment in a randomized complete block design with three replications was conducted in Ghara Chay region, Markazi ...
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To investigate the effects of biological and chemical phosphorus fertilizers, and gibberellic acid (GA3) on yield and its components, and essential oil of Salvia officinalis L., a factorial experiment in a randomized complete block design with three replications was conducted in Ghara Chay region, Markazi province, Iran. Experimental treatments included triple super phosphate at three levels (100, 70, and 30 % of the recommended amount), phosphorus bio-fertilizer of Barvar-2 at two levels (application and no application), and GA3 at three levels (0, 100, and 200 ppm). The results showed that Barvar-2, GA3, and triple super phosphate affected yield and its components, and essential oil significantly. The Barvar-2 application increased the yield and essential oil percentage by 7.7 and 18.2% compared to the control, respectively. It also resulted in a slight increase of the major essential oil compounds. Also, the GA3 200 ppm application increased the percentage and essential oil yield by 23.5 and 40.5% compared to the control, respectively. The highest biological yield and essential oil yield were obtained 3751 and 15.52 kg.ha-1, respectivley with the simultaneous application of 100% of the recommended triple super phosphate, Barvar-2, and GA3 200 ppm. The highest α-thujone percentage (27.91%) as the most important essential oil compound was obtained with the simultaneous application of 100% of the recommended triple super phosphate, Barvar-2, and GA3 100 ppm. The combined application of GA3 and Barvar-2 reduced the use of phosphorus chemical fertilizer to achieve better vegetative characteristics, and high yield and essential oil percentage, so that in many traits the simultaneous application of 70% of the recommended triple super phosphate, Barvar-2, and GA3 did not differ significantly from the 100% of the recommended triple super phosphate application. Therefore, with the combined use of GA3 200 ppm and Barvar-2, the amount of phosphorus fertilizer consumption could be reduced by 30% without causing a significant reduction in yield and essential oil of S. officinalis.
Y. Sarrafi; Gh.R. Moradi Robati; M.H. Fatemi
Abstract
The objective of this study was to investigate the antifungal effects of liquid smoke from tobacco waste on three plant pathogenic fungi and identification and measurement of compounds effective on the antifungal properties of liquid smoke. The waste was pyrolyzed in furnace in the absence of oxygen, ...
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The objective of this study was to investigate the antifungal effects of liquid smoke from tobacco waste on three plant pathogenic fungi and identification and measurement of compounds effective on the antifungal properties of liquid smoke. The waste was pyrolyzed in furnace in the absence of oxygen, and smoke was converted to liquid smoke by refrigerant. Antifungal effects of liquid smoke were tested against Sclerotinia sclerotiorum, Rhizoctonia solani and Phytophthora nicotianae causing tobacco sore shin, collar rot and black shank disease agents, respectively. Six concentrations of the liquid smoke 0, 250, 500, 1000, 2000 and 4000 ppm were prepared and mixed with culture medium. The results of variance analysis showed that the effect of liquid smoke, concentration and the interaction of liquid smoke on concentration in limiting the fungal growth were significant at 1% probability level. Mean comparison of the effect of liquid smoke on these fungi showed that liquid smoke of 25% midrib + 75% scrap pyrolysis in 4000 ppm concentration had 100 percent mycelial growth control on the above fungi. In addition, the liquid smoke pyrolysis of above treatment in low concentrations caused 100 percent mycelia growth control of Rhizoctonia and Sclerotinia. According to research reports, the presence of phenolic compounds, organic acids and alkaloids constituents in liquid smoke is an agent of antifungal effects. Therefore, extraction of these constituents from liquid smoke was carried out and qualitative analysis was done by gas chromatography and mass spectrometry (GC/MS) and quantitative analysis of total phenol and alkaloids was done by spectrophotometer method. The results of this research showed that the effects of alkaloids constituents on the control of fungi were more than phenolic and organic acids constituents in liquid smoke from tobacco waste. The results showed the high potential of the liquid smoke of pyrolysis to be used as a biological control of pathogenic fungi.
S. Anbarestani; A.R. Rezazadeh; A. Rezaei
Abstract
Black cumin (Nigella sativa L.) is a medicinal plant belonging to the Ranunculaceae family. The seeds of Nigella sativa, also known as black seed, are used in traditional medicine as a natural remedy for several illnesses including asthma, inflammation, diabetes, and hypertension. The cell culture of ...
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Black cumin (Nigella sativa L.) is a medicinal plant belonging to the Ranunculaceae family. The seeds of Nigella sativa, also known as black seed, are used in traditional medicine as a natural remedy for several illnesses including asthma, inflammation, diabetes, and hypertension. The cell culture of this plant is important because of its active ingredients and significance in medicine. In this research, the effects of fungal elicitor (zero, 0.5 and 1 mg/L), and sucrose (30, 45 and 60 g/L) on the cell culture of black cumin was investigated as a factorial experiment in a completely randomized design with three replications. The cell growth characteristics and biochemical traits were assayed. The results showed that fungal elicitor and sucrose increased the membrane lipid peroxidation, protein concentration, phenol contents, hydrogen peroxide, and anthocyanin whereas peroxidase activity was significantly decreased compared to control. Generally, sucrose and fungal elicitor increased the growth and activity of the cell and increased the protein production by increasing oxidative stress in cells and increasing the entry of substances into the cell and stimulating metabolism.
F. Ahmadi; M. Modarresi; M.A. Kohanmoo
Abstract
Despite the importance of the German chamomile (Matricaria chamomilla L.), as one of the most widely used medicinal plants in the world, there is little information about the genetic variation within and between its populations in Iran. This study was conducted to investigate the genetic diversity of ...
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Despite the importance of the German chamomile (Matricaria chamomilla L.), as one of the most widely used medicinal plants in the world, there is little information about the genetic variation within and between its populations in Iran. This study was conducted to investigate the genetic diversity of 15 chamomile populations based on morphological characteristics in a randomized complete block design with three replications. Height, flower diameter, receptacle diameter, fresh flower weight, dry flower weight, wet biomass, dry biomass, flowering index, essential oil percentage and Chamazulene percentage were analyzed. The populations had a significant difference for all traits (P≤0.01). The phenotypic and genotypic variation was obtained to be more than 60% for dry weight, dry mass and Chamazulene content. Significant values were recorded for the heritability of the traits including percentage of kemazolen (94.04), the wet biomass (93.93), plant height (90.59), flower diameter (86.52), dry flower weight (84.15), and dry biomass (83.08). The highest and lowest yield (kg/ha) were related to Hamedan (9700) and Eurea (753.33) populations, respectively. The highest percentage of Chamazulene and essential oil content were obtained from Dehroud (14.10) and Jam-o-Riz (0.61), respectively. The flower diameter had the highest correlation (0.62) with the percentage of essential oil. Regression analysis for essential oil percentage as the dependent variable was performed and four traits including flower diameter, height, and fresh weight of flower were introduced into the model, explaining 59% of the variation. In the path analysis for essential oil percentage as the dependent variable, the highest direct effect was related to the fresh weight of flower (1.09) and the highest indirect effect was related to the fresh weight of flower (1.06). The cluster analysis grouped the populations into three groups that did not follow geographical distribution.
I. Bernousi; M. Jafari; J. Ahmadi Dizaji
Abstract
This research was aimed to investigate the effect of two infection methods (immersion and spray) and two Agrobacterium rhizogenes strains (strain A13 and GMI9534) to induce hairy roots on different explants (hypocotyl, cotyledon, leaf, and stem node) of Teucrium chamaedrys. Strain GMI9534 could not induce ...
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This research was aimed to investigate the effect of two infection methods (immersion and spray) and two Agrobacterium rhizogenes strains (strain A13 and GMI9534) to induce hairy roots on different explants (hypocotyl, cotyledon, leaf, and stem node) of Teucrium chamaedrys. Strain GMI9534 could not induce hairy roots in any of the explants, whereas strain A13 was only able to induce hairy roots in leaf and stem node explants. Infection by immersion method was more successful, with higher root induction efficiency (more than 73.3%), and leaf explants showed the highest induction frequency (83.3%). The transformed hairy root lines were confirmed by PCR using rolA and rolB gene-specific primers. Significant differences were shown among the 20 independent hairy root lines cultured on growth regulator-free MS solid medium for total root length and for the root branching. These variables were stable across subcultures and hence seven independent hairy root lines were selected based on these growth properties. Subsequently, the cultures for these hairy root lines were established in half-strength MS liquid medium to monitor their biomass production during a 90-day culture period. Considerable variations were observed in growth capacity among the lines. Line TC-HR-16 produced the highest fresh biomass (9100 mg/30 ml culture medium), a 455-fold increase over initial inoculum, during the 10-weeks culture period. The best-characterized hairy root lines, resulted in this study, can be used to improve the production of secondary metabolites of pharmaceutical values of T. chamaedrys.
A. Pirzad; M. Alizadeh; A. Hassanzadeh Gorttapeh; R. Darvishzadeh
Abstract
To evaluate the effect of seed treatment with gamma rays and nitrogen application on yield and nitrogen use efficiency for protein production in Chamomilla recutita L. leaves, a factorial experiment was conducted based on randomized complete blocks design with three replications. Treatments were gamma ...
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To evaluate the effect of seed treatment with gamma rays and nitrogen application on yield and nitrogen use efficiency for protein production in Chamomilla recutita L. leaves, a factorial experiment was conducted based on randomized complete blocks design with three replications. Treatments were gamma rays from Cobalt-60 source on chamomile seeds cv. Bodegold (0, 4, 8, 12, 16, and 20 Gray) and different levels of nitrogen (0, 50, 100, and 150 kg/ha). Results showed the significant interaction effect of gamma and nitrogen on the leaf yield, biomass and protein, harvest index of protein, and nitrogen use efficiency for leaf and protein production, the yield of dried flower and essential oil. But gamma irradiation and nitrogen had no significant effect on leaf protein and essential oil percentage. The highest yield of leaf (4194 kg/ha) and biomass (19996 kg/ha) were obtained at 8 GRAY of gamma rays with 100 and 150kg/ha of nitrogen application, respectively. However, the highest yield of protein (571kg/ha) was obtained from 20 GRAY of gamma rays and 0 kg/ha of nitrogen and the lowest yield (151kg/ha) was obtained from 0 GRAY and 50kg/ha of nitrogen. The highest value of nitrogen use efficiency for leaf (67.39) and protein (9.30) production was related to the plants grown from seeds and treated by 16 GRAY of gamma and 50kg/ha nitrogen. The lowest nitrogen use efficiency for leaf (13.94) and protein (1.42) production were observed at 16 GRAY and 150kg/ha nitrogen. The highest yield of dried flower in 0, 4, 8, 12, 16 and 20 GRAY were obtained at 150, 100, 0, 150, 100 and 100 kg/ha of nitrogen application, respectively. Generally, irradiations over the 12 GRAY resulted in reducing the nitrogen application for protein production.
M.J. Seghatoleslami; S.G Mosavi; T. Barzegaran
Abstract
Nowadays, changes in cultivation pattern towards drought resistant species are proposed as a solution to tackle drought. To study the effect of irrigation levels and planting date on yield and water use efficiency of Hibiscus sabdariffa L., an experiment was conducted in the research field of agriculture ...
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Nowadays, changes in cultivation pattern towards drought resistant species are proposed as a solution to tackle drought. To study the effect of irrigation levels and planting date on yield and water use efficiency of Hibiscus sabdariffa L., an experiment was conducted in the research field of agriculture college of Islamic Azad University of Birjand during 2010-2011. Three levels of irrigation (20, 60 and 100 percent evapotranspiration of source plant) as main plots and different planting dates (May 10th, June 1th and June 20th) as sub plots were studied in a split plot experiment with three replications in a randomized complete blocks design. The results indicated that different irrigation levels and planting dates had significant effects on number of flowers per square meter, dry and fresh weight of flowers, total biomass yield and water use efficiency (flower and biomass). The highest number of flowers per square meter (62.42), flower fresh weight (62.50 grams per square meter), flower dry weight (10.30 grams per square meter), total biomass yield (150.3 grams per square meter), water use efficiency of flowers (0.024 gram per) and water use efficiency of biomass (0.532 gram per liter) were obtained from the first planting date i.e. May 10th. Also, the highest number of flowers per square meter (55.88), flower fresh weight (72.40 grams per square meter), flower dry weight (11.46 grams per square meter), biomass yield (142.5 grams per square meter), water use efficiency of flower (0.042 gram per liter) and water use efficiency of biomass (0.897 gram per liter) were recorded for the irrigation level of 20 percent evapotranspiration of the source plant. Irrigation levels and planting dates had no significant effect on flower harvest index. The interaction effect of irrigation levels and planting dates was significant on flower dry and fresh weight, total biomass yield and water use efficiency (flower and biomass). The highest values of these traits were obtained at planting date of 10 May and irrigation level of 20 percent evapotranspiration of source plant. Generally, with regard to the early cold fall and long growth period of Hibiscus sabdariffa, its plantation is not recommended in Birjand climate. According to the results of this experiment, water requirement of this plant was low.
R. Rabei; J. Sohrabipour
Volume 23, Issue 1 , May 2007, , Pages 23-38
Abstract
Gracilaria salicornia (C.Agardh) Dawson is an agarophytic alga which has wide distribution in tropical seas. The species has formed small communities in northeast of the Qeshm island in south of Iran. In this study seasonal changes of biomass and two communities of the species were studied since October ...
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Gracilaria salicornia (C.Agardh) Dawson is an agarophytic alga which has wide distribution in tropical seas. The species has formed small communities in northeast of the Qeshm island in south of Iran. In this study seasonal changes of biomass and two communities of the species were studied since October 2001 for a year. The transect–quadrate method used in this study and monthly production and percentage cover of the species were measured in different depths of the intertidal regions of the two communities. In this study Analysis of Variance (ANOVA) revealed that there are significant differences in percentage cover and production (dry weight) of Gracilaria salicornia with respect to depth ranges (shore elevation) (P<0.01). The highest dry production (598.9±67.2 g/m2) and percentage cover (% 69.12±3.7) of Gracilaria salicornia were determined between 2.7 to 3.5 m. depth of sea water.