Agriculture and horticulture
Mohammad Haghaninia; abdollah javanmard; Zahra Ghorbani; mohammad reza morshedloo; Karim Kakaei
Abstract
Background and Objective: In contemporary times, the importance of medicinal plants and their identification in advancing national, regional, and global objectives, such as achieving health, pharmaceutical self-sufficiency, fostering employment, and promoting economic development, ...
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Background and Objective: In contemporary times, the importance of medicinal plants and their identification in advancing national, regional, and global objectives, such as achieving health, pharmaceutical self-sufficiency, fostering employment, and promoting economic development, is indisputable. Mentha suaveolens × Mentha piperita, commonly referred to as grapefruit mint, is recognized as a valuable medicinal plant within the Lamiaceae family. Drought stands out as a major limiting factor for global plant growth, constituting the most prevalent environmental stressor that disrupts the balance between reactive oxygen species and antioxidant defense mechanisms, thereby inducing oxidative stress. Nanotechnology emerges as a modern approach with notable potential to enhance plant resilience against drought stress. Among various nanomaterials, carbon quantum dots, discovered in 2004, possess distinctive characteristics, including dimensions below 10 nanometers, reduced toxicity, and superior biocompatibility compared to metal-based alternatives, owing to their carbonaceous structure. Due to these unique attributes, carbon quantum dots have attracted significant research interest in recent years. Therefore, this study aimed to evaluate the effects of carbon quantum dots on the growth, yield, and essential oil composition of grapefruit mint (Mentha suaveolens × piperita) under drought stress conditions.Methodology: This research was conducted as a factorial experiment in a completely randomized design with four replications in pots during the spring and summer of 2023 at the research greenhouse of Maragheh University, East Azerbaijan, Iran. The first factor was irrigation at 90% of field capacity (FC) as control (normal irrigation), irrigation at 70% FC (moderate stress), and irrigation at 50% FC (severe stress). The second factor consisted of different levels of carbon quantum dots (0, 5, 10, 15, and 20 parts per million). Data collected from sampling were analyzed using SAS statistical software, and means were compared using the least significant difference (LSD) test at the 5% level.Results: Analysis of variance indicated that the effects of different irrigation regimes, fertilizer treatments, and their interaction on all measured traits were significant. The findings revealed that the application of 15 ppm carbon quantum dots under normal irrigation increased fresh weight, dry weight, chlorophyll a, and chlorophyll b contents by 110%, 86%, 184%, and 163%, respectively, compared to no CQD application under severe drought stress. Under moderate stress, the use of 15 ppm carbon quantum dots increased essential oil content by 135% and essential oil yield by 298% compared to severe stress without CQD. Furthermore, the highest linalyl acetate content was obtained with 15 ppm carbon quantum dots under 50% FC, showing a 50.1% increase compared to the control treatment. Similarly, the highest linalool content was recorded under normal irrigation with 10 ppm carbon quantum dots, representing a 67.4% increase compared to no CQD application under 50% FC. Moreover, under normal irrigation, 15 ppm carbon quantum dots reduced malondialdehyde and hydrogen peroxide contents by 51% and 58%, respectively, compared to no CQD application under severe stress. Additionally, under mild stress, 15 ppm carbon quantum dots enhanced the activity of superoxide dismutase, guaiacol peroxidase, and ascorbate peroxidase enzymes by 123%, 111%, and 72%, respectively, compared to no fertilizer application under normal conditions.Conclusion: The results indicate that applying 15 ppm carbon quantum dots can promote growth by modulating secondary metabolites and strengthening defense mechanisms through the activation of antioxidant enzymes and increasing the accumulation of osmolytes such as proline and total soluble sugars, thereby reducing malondialdehyde and hydrogen peroxide levels. This enhancement in grapefruit mint plants contributes to improved drought resilience, ultimately enhancing both the quantity and quality of the essential oil.
Biotechnology
ebrahim dorani; sanaz hemmati asl
Abstract
Background and objectives: Anthraquinones are secondary plant metabolites that play a significant role in the treatment of certain diseases, such as hepatitis and cancer. Anthraquinones are utilized not only in textiles and food production but also in the cosmetics, perfume, and pharmaceutical ...
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Background and objectives: Anthraquinones are secondary plant metabolites that play a significant role in the treatment of certain diseases, such as hepatitis and cancer. Anthraquinones are utilized not only in textiles and food production but also in the cosmetics, perfume, and pharmaceutical industries due to their high antioxidant levels. In vitro culture of plants has provided a suitable alternative method for producing medicinal compounds under controlled conditions, not affected by environmental factors. The hairy root culture system, based on inoculation with Agrobacterium rhizogenes, has become an influential approach for producing secondary metabolites in plant roots, such as ginseng, over the last two decades. One of the advantages of in vitro production of these compounds is the possibility of increasing their yield through the use of influential nutrient elements, as well as inducing the production process and biosynthesis of secondary substances with various biological and non-biological elicitors. The objective of this research was to investigate the effect of salt concentration in the culture medium and sucrose levels on increasing hairy root biomass, and to examine the influence of different abiotic elicitors on enhancing anthraquinone production in madder hairy roots.Methodology: For this purpose, in the first experiment, after inducing hairy roots, the effect of sucrose concentration (30, 45, and 60 g/L) in combination with basal medium salt strength (1/2 MS and MS) on root biomass enhancement was studied in a factorial experiment with a completely randomized design and four replications. In the second experiment, the effects of different elicitors, including titanium dioxide (50 and 100 mg/L), chitosan (100 and 150 mg/L), silver nitrate (50 and 100 mg/L), proline (250 and 500 mg/L), gibberellic acid (1 and 2 mg/L), and naphthalene acetic acid (1 and 2 mg/L), were investigated for their role in increasing anthraquinone (alizarin) production in hairy roots.Results: In the first experiment, the 1/2 MS medium containing 45 g/L sucrose produced the highest biomass, followed by MS containing 45 and 60 g/L sucrose, MS with 30 g/L sucrose, and 1/2 MS containing 60 g/L sucrose. The best medium for biomass production from this experiment was selected for the subsequent experiment, in which the effect of different elicitor treatments for two days on the accumulation of the secondary metabolite (alizarin) was assessed. The results showed that the accumulation of the secondary metabolite in the medium containing 250 mg/L of proline exhibited a significant increase compared to other elicitors, followed by 50 mg/L of silver nitrate and 2 mg/L of gibberellic acid, respectively. The secondary metabolite content in media supplemented with two different levels of naphthalene acetic acid did not differ significantly from that of the control.Conclusion: Overall, this study demonstrated that the optimal root biomass production was achieved in 1/2 MS culture medium containing 45 g/L of sucrose, while the highest anthraquinone content was obtained in the presence of proline.
Biotechnology
M. Sarrafi; M. Mahmoodnia Meimand; M. Dahajipour; M.R. Dehghani
Abstract
Background and objectives: In recent years, Urtica dioica L. has attracted the attention of researchers due to its significant effect on many diseases, especially diabetes. However, more research needs to be done in the field of tissue culture and the effect ...
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Background and objectives: In recent years, Urtica dioica L. has attracted the attention of researchers due to its significant effect on many diseases, especially diabetes. However, more research needs to be done in the field of tissue culture and the effect of elicitors on antioxidant properties and the production of its secondary metabolites. This research aimed to investigate the best hormonal combination and light conditions effective on nettle callogenesis and the effect of methyl jasmonate elicitor on its biochemical properties.
Methodology: For this purpose, using stem explants, the effect of different levels of auxin hormones, naphthalene acetic acid (NAA) (1.5 and 3 mg/l) and 2,4-dichlorophenoxy acetic acid (2,4 D) (0.5 and 1.5 mg/l) and cytokinin hormone, benzylaminopurine (BAP) (0.5 and 1.5 mg/l) were evaluated in two light conditions (light and dark) by measuring fresh weight, dry weight and percentage of callogenesis. The nettle plant's callus growth curve was obtained using liquid culture. After starting the suspension culture, methyl jasmonate treatment was applied with three levels of 1, 0.1 and 0.01 mM, and sampling was done at 24, 48 and 96 hours. The enzyme activity of phenylalanine ammonialyase and polyphenol oxidase and total protein content by extracting in 50 mM Tris-HCl buffer with 8.8 acidity and total phenol, total flavonoid content by extracting in 80% methanol was measured using spectrophotometric and standard methods for control samples and those under different treatments of methyl jasmonate. Also, three metabolites of quercetin, kaempferol and rutin were measured using an HPLC device with a UV detector and 18C column stationary phase. Data from the callogenesis experiment were analyzed in a two-factor experiment in a completely randomized design. The data from the biochemical and HPLC experiments were analyzed in a split-plot in time design, and the means were compared with Duncan's test.
Results: According to the results, the best hormonal combination to produce fresh and dry weight in light conditions includes 1.5 mg/l of NAA and 1.5 mg/l of BAP and in dark conditions, it includes 1.5 mg/l naphthalene acetic acid and 0.5 mg/l of benzyl aminopurine. Also, according to the results, naphthalene acetic acid (62.5%) performed better than 2,4-D (43.5%) in the nettle callogenesis percentage. Lighting was determined as the best light condition for callogenesis in nettle, so the amount of fresh and dry weight and percent of callogenesis in light compared to darkness increased by 4.79, 4.91 and 1.92 times, respectively. There was an increase in polyphenol oxidase enzyme activity, total phenol and total protein in some samples treated with methyl jasmonate compared to the control, and no significant increase was observed in the activity of phenylalanine ammonialyase enzyme and total flavonoid content. According to the HPLC results, the highest amount of rutin metabolite was obtained in 96 hours after the addition of the elicitor and at the concentration of 0.01 mM methyl jasmonate.
Conclusion: The naphthalene acetic acid hormone was more effective than the 2,4-D hormone and light conditions than darkness in nettle callogenesis. Methyl jasmonate elicitor increased the antioxidant property of nettle (48 hours after treatment) by increasing the polyphenol oxidase enzyme activity. However, compared to the control, it had no significant effect on the phenylalanine ammonialyase enzyme activity and total flavonoid. It is suggested that other concentrations of methyl jasmonate be tested to reach optimal conditions for increasing the production of secondary metabolites in nettle in vitro culture conditions.
Agriculture and horticulture
Farhad Bagheri fard sharabiani; Esmaeil Chamani; Mousa Torabi Giglou; Alireza Ghanbari; Younes Pourbeyrami hir
Abstract
Background and objectives: The beneficial effects of the chamomile plant have been attributed to its essential oil, so any factor that affects the quantity of its essential oil will be of interest to researchers. To investigate the effect of plant growth regulators salicylic ...
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Background and objectives: The beneficial effects of the chamomile plant have been attributed to its essential oil, so any factor that affects the quantity of its essential oil will be of interest to researchers. To investigate the effect of plant growth regulators salicylic acid and methyl jasmonate on the production of secondary metabolites in chamomile plants, this experiment was conducted under the native in vitro conditions of Sharbian City.Methodology: To conduct this research, chamomile seeds were collected from Sharbian (N ʺ52 58 37 and E ʺ06 ʹ11 ˚), East Azerbaijan province. After transfer, the seeds were sterilized for 15 minutes in a detergent. Their surfaces were disinfected with 70% alcohol for 45 seconds and then with sodium hypochlorite for 30 minutes. After surface disinfection, they were washed twice with distilled water. The basic culture medium in this study was the MS culture medium. All cultures were placed in the growth chamber at a temperature of 24±2°C and 16 hours of light and 8 hours of darkness. A factorial experiment was performed in a complete random design with three replications. Test treatments included salicylic acid (SA) and methyl jasmonate (MeJA) at five levels (0, 50, 100, 200, 400 μM). Minguez-Mosquera and Perez-Galvez methods were applied to determine photosynthetic pigment amounts. The amount of phenolic compounds in the plant extract was measured with a slight modification based on the Slinkard and Singleton method with the Folin-Ciocalteau reagent. The data related to this research were analyzed using SAS V.9.g.1 statistical software. The comparison of treatment averages was done with Duncan's test at 5% and 1%. Graphs were drawn using Microsoft Excel software.Results: The analysis of variance showed that the interaction effect of the applied treatments on the traits of stem weight, root weight, stem length, and root length is significant at the 1% probability level. In addition, it is significant on the number of stems at the 5% probability level. The comparison of the averages showed that the highest amount of stem weight was obtained in the interaction effect of (50 µM) SA and (200 µM) MeJA, and the lowest amount of stem weight was obtained in the interaction effect of (400 µM) SA and (400 µM) MeJA treatment. The comparison of the means shows a significant difference in this trait. The highest amount of root weight was obtained in the interaction effect of (100 µM) SA and (100 µM) MeJA, and the lowest amount of root weight was obtained in the interaction of (400 µM) SA and (400 µM) MeJA treatment. Came. A comparison of the averages showed that the highest number of stems was obtained in the interaction of (100 µM) SA and (50 µM) MeJA, and the lowest number of stems was obtained in the interaction of (400 µM) SA and (200 µM) MeJA. The comparison of the means showed the significance of the stem and root length traits. The highest value of these two traits was due to the interaction of (50 µM) SA and (100 µM) MeJA treatments, and the lowest value of stem length was in the combination of (400 µM) SA and (µM) treatments. 400) MeJA and root length were measured from the combination of SA (400 µM) and MeJA (200 µM) treatments. In the biochemical traits, variance analysis of the data showed that the effect of SA levels on the amount of chlorophyll a, b, carotenoid, total chlorophyll, phenol, and flavonoid in all three wavelengths was significant at the 1% probability level.Conclusion: In this experiment, it was shown that metabolite production depends not only on enzyme activity but also on stimulant concentration. Increasing the stimulus more than usual not only increases metabolism but also reduces or stops the production of metabolites through the reduction of enzyme activity (probably through the reduction of the corresponding gene expression).
Improvement and breeding
Mostafa Sarani Mallak; Maryam Allahdou; Leila Mehravaran; Halimeh Piri
Abstract
Background and objectives: Drought stress is the most critical factor limiting agricultural and medicinal plants' performance in arid and semi-arid areas. Silybum Marianum L. is a medicinal plant with antioxidant properties. In addition to the plant's genetic nature, flavonolignan ...
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Background and objectives: Drought stress is the most critical factor limiting agricultural and medicinal plants' performance in arid and semi-arid areas. Silybum Marianum L. is a medicinal plant with antioxidant properties. In addition to the plant's genetic nature, flavonolignan production and accumulation are affected by various environmental conditions. The accumulation of secondary metabolites under drought stress was studied concerning the antioxidant defense system at the biochemical level. The purpose is to evaluate the secondary metabolites of milk thistle under non-stress conditions and different levels of drought stress and different growth conditions, as well as to identify the best level of moisture stress and the time of harvesting the plant to increase the effective compounds.Methodology: Milk thistle seeds were disinfected and transferred to a Petri dish containing filter paper and placed in a germinator at 25°C for germination. The germinated seeds were transferred to the pots and put under controlled temperature and humidity in the greenhouse of Hirmand city, Shandel village, located 25 km from Zabul city, Sistan, and Baluchistan province. Evaluation of the effect of drought stress at four different levels of irrigation (25, 50, 75, and 100% of water requirement respectively severe stress, moderate stress, mild stress, and non-stress) and in 3 growth stages (6, 13 and 20 weeks after planting) on biochemical traits including proline content (PC), carbohydrates content (CC), total phenol content (TPC), total flavonoid content (TFC), antioxidant activity and activity of antioxidant enzymes such as catalase (CA), ascorbate peroxidase (AP), guaiacol peroxidase (GP), superoxide dismutase (SOD) and polyphenol oxidase (PO) was carried out. The experiment was done as a factorial based on a completely randomized design with three replications. Data and errors were examined for normality. After confirming the normality of the data and errors, analysis of the variance of the traits and comparing the mean of the traits (LSR) was done at the 5% level.Results: The variance analysis of traits showed that the effect of different levels of irrigation, harvest time, and their interaction on all traits was significant. Comparison of the average interaction effect of irrigation treatment and harvest time of traits: proline content, carbohydrates content, phenol and flavonoid content, and antioxidant activity increased in all growth stages and the lowest and highest values were respectively observed in the growth stage 6 weeks after planting in 100 Percentage of water requirement and growth stage 20 weeks after harvesting in the condition of 25% water requirement. Therefore, the drought stress factor can be used to improve the effective substances of this plant. In addition, the final growth stage is the most appropriate time to harvest this plant due to the accumulation of secondary metabolites at this stage. The interaction effect of irrigation treatment and harvest time was not significant for the activity of guaiacol peroxidase enzyme, and for other antioxidant enzymes it showed that the highest activity of catalase enzyme was at the growth stage 6 weeks after planting in conditions of 25 and 50% water requirement, for ascorbate peroxidase enzyme, it belonged to the growth stage 6 weeks after planting in the condition of 100% water requirement, and for polyphenol oxidase and superoxide dismutase enzymes, it belonged to the growth stage 20 weeks after planting in the condition of 25% water requirement. These results indicate that antioxidant enzymes act differently at different growth stages and under various moisture stress conditions.Conclusion: The evaluation results of milk thistle in 4 irrigation regimes and three growth stages showed that most biochemical traits increased under stress conditions. This indicates that the milk thistle plant responds to drought stress through an enzymatic and non-enzymatic antioxidant defense system. Milk thistle plants had the highest total phenolic and flavonoid content at the final development stage (20 weeks after planting). Therefore, the best time to harvest is at the final stage of development, which has the most polyphenolic compounds.
Improvement and breeding
A.A. Alizadeh Everi; M. Parsaeian; Z. Ghasimi Hagh
Abstract
Glycyrrhiza glabra L. is one of the valuable medicinal plants of the world, whose glycyrrhizin, the most important triterpenoid metabolite in its roots, is widely used in food and modern pharmaceutical industries. To overcome the seed germination problems, the risk of plant extinction due to the indiscriminate ...
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Glycyrrhiza glabra L. is one of the valuable medicinal plants of the world, whose glycyrrhizin, the most important triterpenoid metabolite in its roots, is widely used in food and modern pharmaceutical industries. To overcome the seed germination problems, the risk of plant extinction due to the indiscriminate root harvesting, and the slow rate of natural biosynthesis of secondary metabolites, it is necessary to study on this plant root tissue culture and use elicitors to increase its glycyrrhizin biosynthesis. In the present study, the seed germination rate of two Iranian licorice ecotypes (Kashmar and Joghtai) and an Iraqi ecotype was investigated at different levels of sulfuric acid pretreatment. In addition, differences between the ecotypes in terms of morphology and root glycyrrhizin content were studied in response to IAA and NAA auxin elicitors with concentrations of 0, 0.5, 1, and 1.5 mg.l-1 in ½ MS medium using root explants. The results showed that all three ecotypes obtained the highest seed germination in the 98% sulfuric acid pretreatment for 40 minutes. The Iraqi ecotype was placed in the statistically best group in terms of all traits except glycyrrhizin content under control and some hormone levels (esp. NAA) conditions. In the Iranian ecotypes, the auxin elicitors application significantly improved the root traits and glycyrrhizin content. Kashmar ecotype produced the thickest roots using 0.5 mg.l-1 of IAA and the highest root dry weight (5.8 and 5.4 mg, respectively) using the medium and high concentrations of NAA and IAA. Joghtai ecotype also produced the heavy roots with 1.5 and 0.5 mg.l-1 of IAA and NAA (100 and 79 mg, respectively). In general, the medium and high concentrations of IAA and NAA (1 and 1.5 mg.l-1) caused the highest glycyrrhizin production (8.82 and 7.83 µg.g-1 DW, respectively) in Kashmar ecotype roots. Therefore, selection of appropriate ecotype and auxin elicitors application can increase in vitro production of biomass and root glycyrrhizin content in G. glabra.
Sh. Gheidarlouei; R. Khademian; S. Mafakheri
Abstract
To investigate the effect of mycorrhiza inoculation on increasing the resistance of medicinal plant dill (Anethum graveolens L.) to salinity stress, a factorial experiment was conducted in a randomized complete block design with three replications at the research greenhouse of Imam Khomeini International ...
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To investigate the effect of mycorrhiza inoculation on increasing the resistance of medicinal plant dill (Anethum graveolens L.) to salinity stress, a factorial experiment was conducted in a randomized complete block design with three replications at the research greenhouse of Imam Khomeini International University, Qazvin province, Iran. The first factor was ecotype at two levels (Isfahan and Varamin), the second factor was salinity at three levels (0, 5 and 10 dS/m), and the third factor was mycorrhiza at three levels (0, 75 and 150 g fungi). The results showed that mycorrhizal inoculation improved all the quantitative traits studied so that under salinity conditions, the plants inoculated with mycorrhiza showed better growth than non-inoculated plants. The highest dry weight, plant height, number of seeds plant-1, and 1000-seed weight were obtained in ecotype Isfahan, salinity level 5 dS/m and 150 g of mycorrhiza. Eleven compounds were identified in dill essential oil. The highest percentage of limonene (7.5%) was obtained from ecotype Varamin plants treated with 5 dS/m salinity and 150 g mycorrhiza. The highest percentage of carvone (86.3%) was obtained in ecotype Isfahan treated with 150 g mycorrhiza and non-salinity.
R. Hazrati Jahan; N. Zare; S. Dezhsetan; P. Sheikhzadeh Mosaddeg
Abstract
The discovery of taxanes production in hazelnut (Corylus avellana L.) and its cell cultures has generated considerable interest and hopes for studying and utilization of cell suspension culture of hazelnut as a biotechnological approach to the taxol production. In this research, the effects of ...
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The discovery of taxanes production in hazelnut (Corylus avellana L.) and its cell cultures has generated considerable interest and hopes for studying and utilization of cell suspension culture of hazelnut as a biotechnological approach to the taxol production. In this research, the effects of plant growth regulators (different levels of 2,4-D, NAA, Kin and BAP) on hazelnut cell culture growth, and effects of elicitor (methyl jasmonate, chitosan and ultrasonund) and precursor (aminobenzoic acid and phenylalanine) combination on taxol production in cell suspension cultures were investigated. The results revealed that hazelnut cell growth in suspension cultures was significantly influenced by growth regulators treatments. Cell growth in MS medium containing 1 mg/l 2,4-D, 0.5 mg/l BAP and 150 mg/l ascorbic acid was significantly higher than other treatments. The cell growth and viability, EC and pH of medium and taxol production were significantly different between elicitor treatments. Overall, elicitor treatments inhibited cell proliferation but significantly increased the taxol production compared to the control (without elicitor). The highest taxol content (15.27 mg/l) was obtained with 2 min US and 20 mg/l aminobenzoic acid treatment in the cells derived from seed explants, which was about 132.78- fold of control.
F. Ahmadi; A. Moieni; S. Rashidi Monfared
Abstract
In this research, Echinacea purpurea L. hairy roots were cultured in 1000 ml bubble column bioreactor containing 500 ml WPM liquid medium and the effects of inoculum densities (3, 6 and 9g l-1) and aeration rates (0.1, 0.2 and 0.4vvm) on biomass and cichoric acid production were investigated. Fresh and ...
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In this research, Echinacea purpurea L. hairy roots were cultured in 1000 ml bubble column bioreactor containing 500 ml WPM liquid medium and the effects of inoculum densities (3, 6 and 9g l-1) and aeration rates (0.1, 0.2 and 0.4vvm) on biomass and cichoric acid production were investigated. Fresh and dry weights and also the cichoric acid content in hairy roots were measured after 30 days. The results showed that the highest amount of biomass (16.4g l-1 fresh weight and 2.084g l-1 dry weight) and cichoric acid (16.74mg g-1 DW) were produced in the inoculum density of 6 gl-1. The investigation of aeration rates on biomass and cichoric acid production showed that the maximum fresh weight (15.4g l-1) and dry weight (2.467g l-1) were obtained in aeration rate of 0.4vvm and the maximum cichoric acid content (12.74mg g-1 DW) was obtained in aeration rate of 0.2vvm. Overall, the inoculum density and aeration rate had considerable effects on the hairy root growth and development in bubble column bioreactor and they should be optimized for obtaining the highest hairy root biomass and secondary metabolites.
