Medicinal Plants
Alireza Moshrefi-Araghi; Seyyed Hossein Nemati; Mahmood Shoor; Majid Azizi; Nasrin Moshtaghi
Abstract
Introduction Mentha is one of the most important genera of aromatic plants which belongs to the Lamiaceae family. The genus of Mentha is distributed across Asia, Africa, Australia, Europe, and North America. Mentha longifolia L., also known as wild mint, is a fast-growing aromatic perennial herb. ...
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Introduction Mentha is one of the most important genera of aromatic plants which belongs to the Lamiaceae family. The genus of Mentha is distributed across Asia, Africa, Australia, Europe, and North America. Mentha longifolia L., also known as wild mint, is a fast-growing aromatic perennial herb. It is widely used as herbal medicine and is beneficial for the immune system and fighting with secondary infections. The essential oil of this plant is partly responsible for the decongestant, antispasmodic and antibiotic effects. Currently, much emphasis is being laid on conserving plant germplasm as valuable bio-resources. Selection between and within accessions for a high level of herbage yield and other characters requires an effective tool to be applied by mint breeders. Achieving to cultivars that are more capable of optimum producing is a breeding goal. The objectives of this study were to analyze the diversion of the agronomical traits of Mentha longifolia L. genotypes from different regions of Iran under a similar condition in order to find the superior genotypes and introduce for the domestication of this plant. Material and MethodThis research was performed at the research field of the Ferdowsi University of Mashhad. The field is located at 36˚15' North latitude and 59˚38' East longitude, at an altitude of 985 meters. The information related to temperature and precipitation was obtained from climate station. Soil sampling was done in the depth of 0.3 meter and physical experiments on samples were done before starting the experiment. The field was fertilized by 25 kg/ha animal manure. Seeds of 20 genotypes of M. longifolia L. were prepared from Gene Bank of Research Institute of Forests and Rangelands and a view of the wild mint genotypes distribution was presented on the map. The experiment was performed in a field with 500m2 areas in 2015-2016 growing seasons. The experiment in form of compound analysis arranged in complete randomized design with three replicates of 20 wildmint's genotypes in every replication. In each furrow, fourteen bush was totally studied, in which the distance between the two plants was 20 cm. The plot was considered 1.2 by 3 m and the distance between rows was 0.50 m. The dimension in the plots for every replication area was 3.6 m2 and the distance between blocks were 1.5 and between experimental units were 0.5 meters. Picking up was done after elimination of 0.5 meters from every side of replication. The essential oil was extracted using Clevenger type apparatus and by hydrodistillation. Results and DiscussionAnalysis of variance showed that the effect of the diversity among the genotypes on morphological, agronomical and essential oil yield parameters of wild mint was significant (P≤0.01). The results of this study showed that Mentha longifolia L. herb has a high diversity under the same culture conditions among genotypes collected from 20 regions of Iran. Correlation result shows that vegetative organs have more effect on the essential oil content than the reproductive organs. This may be due to the distribution of essential oil accumulation and storage sites, which requires further research in this regard. The study of vegetative and reproductive characteristics suggests that the genotypes of the dry and semi-arid climates have different conditions, which may causes the separation of their genotypes compared with the other genotypes. The bi-plot, based on PC1 and PC2, reflects the relationships between the studied genotypes. The results of bi-plot of cluster analysis confirmed that genotypes were divided into two main groups based on agronomical and morphological traits. Conclusion Finally, plants of genotype G13 belongs to Hormozgan province and then G16 genotype belongs to Khuzestan province were selected because of more biomass, more aerial part volume and also the most essential oil performance compared to the other genotypes. Desirable traits for the aerial part such as the time of flowering, leaf length, leaf width, plant height, the highest number of leaves in the stem and internode distance are important traits that should be considered. In addition, the cultivation of these genotype in order to the further production can be the great help in the domestication of this species and, given the fact that the diversity is a precursor of breeding, this research can be an introduction for future breeding operations. However, further research is needed to confirm the phytochemical superior genotypes.
M. Zare Hasan Abadi; A. Ganjeali; M. Lahouti; N. Moshtaghi
Abstract
Introduction: Agrobacterium rhizogenes hairy roots induction is used for secondary metabolite production in plants. A. rhizogenes is a genus of gram-negative soil bacteria belonging to the Rhizobiaceae family that causes hairy roots at the site of infection. Hairy roots have various advantages, including ...
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Introduction: Agrobacterium rhizogenes hairy roots induction is used for secondary metabolite production in plants. A. rhizogenes is a genus of gram-negative soil bacteria belonging to the Rhizobiaceae family that causes hairy roots at the site of infection. Hairy roots have various advantages, including high growth rate, more genetic stability than the callus and suspension cultures, growing well on hormone-free media that have been reported effective for producing high levels of secondary metabolites. Basil (Ocimum basilicum) is a popular herb with important economical applications in food, cosmetic, and pharmaceutical industry. It is a digestive stimulant with anticarcinogenic, antibacterial, and anticonvulsant properties. The main phenolics reported in basil plants are in the classes of phenolic acids and flavonoids, some of which have human health benefits. This study was designed to develop hairy root culture from O. basilicum using different of A. rhizogenes strains for the production of total phenols and introduce the best strain of A. rhizogenes to induce hairy root and growth and production of total phenol.
Materials and Methods: Different A. rhizogenes strains (ATCC-15834, A4, MSU, and R1000) were studied to investigate their effects for the transformation and production of secondary metabolites in O. basilicum. Therefore, shoot and leaf explants and nodes of the seedlings were used for Agrobacterium-mediated transformation. These explants were inoculated with four A. rhizogenes strains and transferred to ½ MS medium. About four weeks after cultivation with A. rhizogenes, hairy roots were excised from the seedlings and subcultured to fresh medium MS liquid culture containing 500 mg/l cefotaxime. After 60 days of inoculation, various parameters, including dry weight, infection percentage, number of hairy roots per explant, and total phenol contents were measured. The growth rate and phenolic contents of the transformed hairy roots were compared with normal ones. Total genomic DNA was isolated from non-transgenic and transgenic hairy root lines using the Cetyl Trimethyl Ammonium Bromide (CTAB) method. Isolated genomic DNA was used to detect the rolC gene through polymerase chain reaction (PCR) analysis. The PCR using specific primers for rolC of T-DNA and virD2 was used to confirm the nature of resulted transgenic hairy roots.
Results and Discussion: Selecting efficient A. rhizogenes strains, as well as the type of explants, are crucial factors for hairy root induction. All used A. rhizogenes strains were able to produce hairy roots. Hairy roots appeared on the nodes at the point of injection, but were not forming on the shoot and leaf explants. So, the choice of the plant material is crucial for successful transformation with A. rhizogenes and usually, transformation of young tissues gives the best results. The transgenic status of the hairy roots was confirmed using PCR with rolC and virD specific forward and reverse primers. All lines showed the presence of 612 bp rolC amplified products, indicating the integration of T-DNA of A. rhizogenes and O. basilicum. Hairy roots could synthesize phenolic compounds, which was significantly higher in hairy roots than non-transformed control. Four hairy root lines were independently evaluated for their content and these lines showed variation in total phenolic contents, with the highest amount (312 mgGAE/ g DW) in hairy roots induced by ATCC-15834 strain and the lowest amount (113.2 mgGAE/ g DW) in hairy roots induced by R1000 strain. The results showed that the strain ATCC-15834 caused the highest infection percentage (68.1%) along with the highest number of hairy roots (4.8) per explant and root length (1.8 cm). The growth rate and phenolics production were investigated in each hairy root of O. basilicum from infection by four different A. rhizogenes strains. The highest growth rate (103.2 mg DW) and production of total phenol (312 mg/g DW) were found in ATCC-15834. The growth rate of transformed hairy roots was more than that of normal ones.Total phenol contents in all hairy roots were also increased significantly compared with non-transformed control plants (4.6 times in hairy roots induced by A. rhizogenes strain ATCC-15834). ATCC-15834 has been reported as the most widely used A. rhizogenes strain owing to its strong induction ability, and the variation in hairy root induction could be due to disparity in the virulence of different A. rhizogenes strains.
Conclusion: The hairy roots of O. basilicum had shown promising results in terms of significant yield of phenolic contents and had the potential for being scaled-up further for phenol production. It could be concluded that A. rhizogenes strains had different abilities in hairy roots induction. Therefore, the selection of an effective A. rhizogenes strain for the production of transformed root cultures is important, highly dependent on the plant species, and must be determined in future experiments.
Hoda Zare Mirakabad; Mohammad Farsi; Saeid Malekzadeh Shafaroudi; Mehrdad Iranshahi; Abdolreza Bagheri; Nasrin Moshtaghi
Abstract
Introduction: There is a growing body of the literature that recognizes the importance of ferutinin (C22H30O4) as one of the natural phytoestrogens with potency to treat osteoporosis and some kind of cancers. One of the greatest challenges is availability of ferutinin that is found just in root of some ...
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Introduction: There is a growing body of the literature that recognizes the importance of ferutinin (C22H30O4) as one of the natural phytoestrogens with potency to treat osteoporosis and some kind of cancers. One of the greatest challenges is availability of ferutinin that is found just in root of some plants of the genus Ferula (Apiaceae), which is the reason of high price of it in global markets. Ferula ovina , an endemic plant of Iran, is known as one of the greatest sources of ferutinin. Unfortunately, access to ferutinin requires uprooting Ferula ovina especially older plants with more secondary metabolites content. There is a large volume of published studies describing the importance of tissue culture in propagation of endangered plants including secondary metabolites without possibility of chemical production and with deep dormancy seed exactly like the characters of F. ovina. Up to now, far too little attention has been paid to importance of tissue culture in accessing ferutinin without degrading the germplasm resources of it. The main aim of this study was to find an approach to access ferutinin associated F. ovina germplasm conservation.
Materials and Methods: In this experiment, the aerial parts of F. ovina were collected from Zoshk area (Mashhad, Iran). The plants were recognized as F. ovina by the Institute of Plant Sciences (Ferdowsi University of Mashhad). Tissue culture part was performed with preparation of sterilized media and explants. Therefore, the MS salts and vitamins was applied as basic medium, however MS salt was decreased to half strength for rooting of shoots. The node (root junction) explants were cultured on 24 callus induction/shooting media with different combinations of plant growth regulators (BAP, IAA, Kin, NAA and IBA). The shoots from direct and indirect regeneration were then transferred to media with different combinations of PGRs (BAP, IBA, IAA and NAA) in order to find rooting medium. Following these treatments, unbalanced ANOVA for analysis of data were performed using IBM SPSS 19. The final stage of the study comprised a TLC test for the purpose of finding ferutinin in samples which resulted from tissue culture. For this purpose, air-dried parts of samples were powdered and extraction was done after being in 3-5 times dichloromethane for 24hours. Then after optimizing solvent system was done with selection the ratio that presented bands in middle of TLC paper.
Results and Discussions: The results indicated that from 24 tested media, just 8 of them had potency of callus formation, but just L6 on MS medium containing 1 mg/l BAP and 1.5 mg/l IAA showed significant difference for percentage of callus induction at 5% level with compact green and the heaviest calluses. Although direct and indirect shoot regeneration was observed in this study, L9 on MS medium along with 1.5 mg/l IAA and 3 mg/l kin demonstrated significant difference for percentage of shooting at 5% level. Moreover, R6 on ½MS with 1 mg/l IBA showed significant difference for percentage of rooting of shoots at 5% level. The most surprising observation to emerge from the data comparison was L24 on half strength MS medium containing 0.2 mg/l BAP and 3 mg/l IBA with potency of callus induction, shooting and rooting of shoots. Regarding to the results of TLC test, ferutinin positive bands were observed in some samples.
Conclusions: The aim of the present research was to examine possibility of achieving ferutinin without need to uproot F. ovina because there are several problems to achieve this valuable sesquiterpene: 1) Chemical production of ferutinin is impossible, 2) It could be accessible just from roots of genus Ferula, 3) Propagation with seeds of F. ovina is limited because of morphophysiological dormancy of them, 4) Natural habitat of this plant in Iran is going to be destructed, 5) Access to natural habitat is difficult, 6) Time of access is limited with short growth season, 7) Maintaining F. ovina in greenhouse condition is impossible. The results of this research support the idea that producing ferutinin in Iran without any harmful effect on germplasm resources of F. ovina is possible. This is the first study of high scale commercial production of ferutinin which examine associations between tissue culture and ferutinin production.
saba nejatie zadeh; Saeid Malekzadeh Shafaroudi; A. Astaraei; Nasrin Moshtaghi
Abstract
Introduction: An emerging field of nanotechnology in recent years is the use of nanoparticles and nanomaterials in agricultural systems which is due to their excellent mechanical, electrical, optical, surface properties, crop protection and nano-fertilizers. Titanium dioxide (TiO2) is a class of nanoparticles ...
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Introduction: An emerging field of nanotechnology in recent years is the use of nanoparticles and nanomaterials in agricultural systems which is due to their excellent mechanical, electrical, optical, surface properties, crop protection and nano-fertilizers. Titanium dioxide (TiO2) is a class of nanoparticles which widely used in the food industry, cosmetics, papers, pharmaceuticals, plastics and industrial raw materials. The widespread industrial application of TiO2 is due to its white pigment, ultraviolet blocking property, and chemical features commonly used to alleviate pollutants concentration in water, soil and air. Owing to its increasing use in the industry, a large part of TiO2 residues are released into the environment, and currently, TiO2 nanoparticles are being considered an emerging environmental contaminant. However, there have been a number of studies reporting beneficial effects of TiO2 on growth and physiological traits of crops. It has been postulated that the TiO2-induced improvement of crop growth is not merely related to the promotion of photosynthesis; other biochemical processes especially nitrogen metabolism are also involved in this event. Ethylene diamine tetraacetic acid (EDTA) is a widely used as a chelating agent, i.e., the chemical is able to sequester metal ions such as Ca2+ and Fe3+. EDTA is used as nitrogen source for doping of TiO2 nanoparticles which improves TiO2 photocatalytic features. The present study was conducted to investigate the effects of TiO2 nanoparticles and EDTA on growth indices and biochemical parameters in spinach (Spinacia oleracea). For detailed evaluation of treatment effects, different concentrations of TiO2 nanoparticles were sprayed on spinach leaves and the samples were collected in a time course.
Materials and Methods: A factorial experiment was carried out in the form of completely randomized design (CRD) with three replications. Soil samples were taken before cultivation of spinach (S. oleracea) seeds (Var VIROFLAY) and analyzed for nutrients’ concentration. Treatments include different levels of TiO2 (T1=0, T2=0.05mg/l and T3=0.1mg/l) and two concentrations of EDTA (E1=0 and E2=130mg/l) sprayed on spinach plants in research greenhouse of agriculture faculty, Ferdowsi University of Mashhad. Aqueous solutions of nanoparticles were treated by ultrasound for 10 min to enhance homogeneity. The solutions were sprayed on the plant at six- leaves stage. The plant samples were taken before reproductive phase for measurement of biochemical parameters. Nitrogen content of plant samples was measured by PDV 500 Macro- Kjeldahl device; Potassium content was determined by 310c flame photometer; phosphorus concentration in plant samples was measured by spectrophotometer model 2100. Chlorophyll and carotenoid contents were measured by the method proposed by Lichtenthaler (1978). For analysis of growth parameters, plant samples were taken a week after TiO2 treatments and leaf area, shoot fresh/dry weight, stem length, internode length, root area, root fresh/dry weight and total root diameter were measured.
Results and Discussion: Application of 0.05mg/l of TiO2 nanoparticles without EDTA resulted in 13.5% and 9.48% increase in nitrogen and protein; respectively, however by increasing nanoparticles to 0.1mg/l, nitrogen and protein content in the treated plants were respectively reduced to 21% and 19.57% of those of control group (p
Ahmad Noroozi; Abdolreza Bagheri; Nasrin Moshtaghi; Ahmad Sharifi
Abstract
Introduction: Anthurium is a popular genus of the Araceae (order Spathiflorae).The flower consists of a protruding spadix containing numerous florets, subtended by a brightly colored modified leaf, the spathe. Anthuriums are bisexual and protogynous.Anthuriumscherzerianum as the most important species ...
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Introduction: Anthurium is a popular genus of the Araceae (order Spathiflorae).The flower consists of a protruding spadix containing numerous florets, subtended by a brightly colored modified leaf, the spathe. Anthuriums are bisexual and protogynous.Anthuriumscherzerianum as the most important species ofAnthurium genus is a potted perennial plant. Due to having beautiful, attractive and long-life flowers, A. scherzerianum can be used for the production of pot and cut flowers. Tissue culture is suggested as the most commonly method in order to rapid propagation and removing disease in a short period of time. This method also recommended for Anthuriumbecause of problems in classical propagation method of this flower..The three basic propagation methods for Anthuriumare propagation by seed, traditional vegetative and tissue culture.Micropropagation of Anthurium is using forcommercial production.
Materials and Methods: In this study, the effect of plant growth regulators and explants on indirect regeneration of A. scherzerianumdetermined in separate experiments. In the first experiment, callogenesis was done by leaf explants on MS medium containing growth regulators, BA in three concentrations (0.5, 1.25 and 2 mg/l) in combination with
2, 4-D (0.5, 1.25 and 2 mg/l) or NAA (0.5, 1.25 and 2 mg/l) and the combinations of TDZ (0.5, 1.25 and 2 mg/l) with
2, 4-D (0, 0.5 mg/l). In the second experiment, regeneration was done on MS medium containing 0.75 mg/l BA with 0.05 mg/l 2, 4-D and 0.1 mg/l NAA and also in combination with TDZ (0.75mg/l). For rooting, MS medium containing different concentrations of IBA and IAA (0, 0.2 and 1 mg/l) were used. Callus induction, regeneration and rooting experiments were done based on completely randomized design, with 12, 6 and 6 replications, respectively.Data from all the schemes used in this study were analyzed with SAS statistical software. The comparison of means using Duncan's multiple range test was evaluated at the 5% level.
Results and Discussion: Analysis of variance showed that the effect of explant type and hormone combinations was significant on the percentage of callogenesis, callus volume and survival percentage. The interaction effect of explant type and combination of hormones was also significant on percentage of callogenesisand the volume of callus. Means comparisons showed that the highest callogenesis, viability and callus volume were achieved on MS medium containing 2 mg/l of BA and 0.5 mg/l of 2, 4-D. Petiole explants, also produced the highest percentage of callus (95%), survival rate (96%) and callus with dimensions of 6 mm2. Callus formation in leaf vein explants was higher than others. The effect of explant type and hormone combinations on regeneration, number of branches, number of leaves and leaf length was significant.The interaction of explant and hormone combinations on regeneration, number of branches, number of leaves and leaf length was also significant. Moreover, results of regeneration experiment indicated that the maximum number of shoots (6.9) and the maximum shoot length (5 cm), number of leaves (18) and the leaf length (2.8 mm) were achieved in 0.75 mg/l BA mg/l of and 0.05 mg/l 2, 4-D. In this study, petiole explants were also regenerated earlier than leaf explants.The effect of hormone combinations and concentrations was significant on rooting specially on the number of roots and root length.Furthermore, results of rooting experiment revealed that the highest rooting percentage (95%), the maximum number of roots (4.5 per plantlet) and the longest roots (3.5 cm) were produced in the medium containing 0.2 mg/l of IBA. Finally, the rooted plantlets were adapted (90%) in vivo condition by placing them on a mixture of cocopeat and perlite (2:1) substrate.
Conclusion: In this study callugensis, regeneration and rooting of A.scherzerianum’s petiole and leaf explants were studied and different levels of plant growth regulators used for callugensis and regeneration. In this study petiole explants showed the highest callugenesis and regeneration. MS medium containing BA (2 mg/l) and 2, 4-D (0.5 mg/l), was the best for callugenesis. Also the highest percentage of regeneration was observed in medium containing BA (0.75 mg/l) and 2, 4-D (0.05 mg/l). Moreover low concentration (0.2 mg/l) of auxin has a better effect on rooting than high levels (1mg/l) so that the highest rooting percentage was produced in medium containing IBA (0.2 mg/l) and the lowest rooting percentage was produced in medium containing IAA (1 mg/l). Anthurium plantlets acclimized is cocopeat and perlite substrate (2: 1) with 90% acclimation.
A. Khazaei; N. Moshtaghi; Saeid Malekzadeh Shafaroudi; K. Ghous
Abstract
Introduction: Jujube (Ziziphus jujuba) is one of the most important fruit trees in Asia which has been planted from 3,000 years ago in China for medicinal purposes. Jujube belongs to the Rhamnaceae family. The Jujube fruit is used in fresh and dry forms. The fruit is full of vitamin C and has anticancer ...
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Introduction: Jujube (Ziziphus jujuba) is one of the most important fruit trees in Asia which has been planted from 3,000 years ago in China for medicinal purposes. Jujube belongs to the Rhamnaceae family. The Jujube fruit is used in fresh and dry forms. The fruit is full of vitamin C and has anticancer and medicinal effects. This tree can grow on salty and dry lands in Iran. Therefore, increasing the cultivation area of Jujube can be effective for soil conservation. In the last 20years, cultivation of Jujube is is considerable in Iran specially in the South Khorasan Province and 98 % of total production of Jujube in Iran belongs to this province. The low rate of seed germination and low production of shootlets are the most important problems in Jujube proliferation, so micropropagation of this plant through tissue culture was considered.
Materials and methods: In this study, Cangan ecotype of Jujube was used for multiple shoot regeneration. At the end of May, apical buds of shoots were cut from mature trees of the Research Collection of Jujube at Sarbishe, Birjand, South Khorasan Province in Iran. The buds were disinfected with 70% ethanol for 1 min and 2% sodium hypoclorite for 25 min. Then the buds were rinsed with distilled water for 25 min completely. Apical buds were placed on Murashige and Skoog (MS) medium supplemented with different concentrations of BA (0.5, 1, 1.5, 2 mg/L) in combination with IBA or NAA (0, 0.1, 0.2, 0.4 mg/L). After one month, the shoots with 3-5 cm length were transferred to rooting media (1/2 MS + IBA or IAA : 0.5, 2, 5, 10 mg/L). The data were recorded after shooting and rooting and were analysed in the facorial experiment.
Results and Discussion: The results of variance analysis and mean comparisons showed that there are differences between different levels of IBA and BA alone for the number of shoots and their length (P
Amirghaffar Ghaffar Shahriari; Abdolreza Bagheri; Ahmad Sharifi; Nasrin Moshtaghi
Abstract
Abstract
Alstroemeria is one of the most important cut flowers in the world which is multiplicated by rhizome explants in in vitro conditions . The major problem in using rhizome explants is fungal and bacterial contamination which inhibit the preparation of strill explants. Therefor in this experiment, ...
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Abstract
Alstroemeria is one of the most important cut flowers in the world which is multiplicated by rhizome explants in in vitro conditions . The major problem in using rhizome explants is fungal and bacterial contamination which inhibit the preparation of strill explants. Therefor in this experiment, the effect of disinfecting compounds like sodium hypochlorite, mercury chloride, nano silver particles, the antibiotics of streptomycin, penicillin, cefotaxime and carbendazim fungicide were examined on controlling of contamination. After disinfection, the explants were cultured on MS medium supplemented with 2 mg/L BA and 0.2 mg/L NAA. Contamination percentage was measured after three weeks. The results showed that the treatment of plants with 0.4% carbendazim fungicide and disinfection by 70% ethanol for one minute and %3 sodium hypochlorite for 20 minutes for caralis cultivar, and disinfection by 70% ethanol for one minute and %3 sodium hypochlorite for 20 minutes follow by culturing in medium with 200 mg/l either of streptomycin and penicillin in Bordeaux cultivar had the lowest contamination percentage. Contamination was not controlled by nano silver particles in studied plants.
Keywords: Alstroemeria, sodium hypoclorite, mercury chloride, antibiotics, carbendazim
