Biotechnology
S.M. Mostafavi; M.R. Abdollahi; D. Dastan; H. Sarikhani
Abstract
Caper (Capparis spinosa L.) is a rich source of rutin, plays an essential role in human health. In the present study, the effects of cold (25°C as control, 4°C, and 7°C for 2, 4, and 7 days), heat (25°C as control, 30°C for 14 days, 32°C for 2 and 4 days, and 35°C for 8 hours), ...
Read More
Caper (Capparis spinosa L.) is a rich source of rutin, plays an essential role in human health. In the present study, the effects of cold (25°C as control, 4°C, and 7°C for 2, 4, and 7 days), heat (25°C as control, 30°C for 14 days, 32°C for 2 and 4 days, and 35°C for 8 hours), and carbohydrate treatments on the androgenesis efficiency were studied in the anther culture of caper. Also, the effects of maltose and sucrose at the concentrations of 30 and 60 g L-1 in combination with two temperature treatments (1- 30°C for 14 days and 2- 7°C for 7 days + azacytidine and 2,4-D pretreatments) on the androgenesis induction was evaluated. The temperature and carbohydrate treatments showed statistically significant differences (p < /em>≤0.01) in terms of callus and embryo formation. The 7°C for 2, 4, and 7 days produced the highest percentage (at the third week: 80, 78.34, and 76.67%, respectively) and callogenesis speed (7.85, 7.75, and 7.60 calli week-1, respectively) and the 7°C for 7 days produced the highest embryo production (0.57 embryo anther-1). The 30°C for 14 days treatment showed the highest percentage (at the third week: 100%) and callogenesis speed (9.44 calli week-1). While the 32°C for 2 and 3 days and also 30°C for 14 days produced the highest number of embryos per anther (0.22, 0.20, and 0.18 embryo, respectively). The use of 30 g L-1 maltose in combination with the 30°C for 14 days produced the highest percentage (at the third week: 91.66%) and callogenesis speed (8.94 calli week-1), while the 30 g L-1 maltose in combination with the 7°C for 7 days + azacytidine and 2,4-D pretreatments produced the highest mean embryo number per anther (0.55 embryo). The results of this research are of great importance for the use in the caper breeding programs.
V. Payamnoor; J. Nazari; R. Jafari Hajati
Abstract
Betulin and betulinic acid are from the most important anticancer and anti-HIV metabolites, and the birch species (Betula spp.) bark is considered as the primary source of these metabolites. Due to the extinction of these tree species in Iran, it is necessary to replace the metabolites extraction from ...
Read More
Betulin and betulinic acid are from the most important anticancer and anti-HIV metabolites, and the birch species (Betula spp.) bark is considered as the primary source of these metabolites. Due to the extinction of these tree species in Iran, it is necessary to replace the metabolites extraction from the birch bark with modern methods such as cell and tissue culture to produce the metabolites. The aim of this study was to determine the effect of explant type on the amount of betulin and betulinic acid produced in calli of two birch species B. pendula and B. litwinowii under in vitro conditions compared to the amount of metabolites extracted from the tree bark. Bark and leaf explants of two mentioned species were cultured in WPM medium containing 1 mg/L BAP and 0.1 mg/L 2,4-D for callogenesis. The amount of betulin and betulinic acid in three-month calli was measured using the HPLC technique and compared with the amount of these metabolites in one-centimeter stem bark samples taken from nature. The bark explant was more successful in callogenesis, and calli derived from this explant had more active ingredients. The amount of betulin and betulinic acid from the extract of bark sample taken from nature was respectively obtained to be 5.23 and 2.91 percent for B. pendula, and 5.65 and 2.52 percent for B. litwinowii. Moreover, calli derived from the bark explant of B. pendula and B. litwinowii contained 0.023 and 0.016 percent of betulin and 0.053 and 0.057 percent of betulinic acid, respectively. Generally, the results indicated that the bark explant was more capable of callogenesis and secondary metabolite induction than the leaf explant in both birch species under in vitro conditions.
V. Payamnoor; R. Jafari Hajati
Abstract
Birch (Betula pendula Roth.) is one of the most important species exposed to extinction due to the limited natural habitat and lack of regeneration. Plant tissue culture technique is an appropriate solution for asexual reproduction of this species and to protect it in controlled conditions. In ...
Read More
Birch (Betula pendula Roth.) is one of the most important species exposed to extinction due to the limited natural habitat and lack of regeneration. Plant tissue culture technique is an appropriate solution for asexual reproduction of this species and to protect it in controlled conditions. In this research, callogenesis of B. pendula was investigated using five types of explants (leave, stem, nodal, petiole and bark) in WPM and NT media enriched with two hormonal combinations including A) 1mg/l BAP, 0.1 mg/l 2,4-D and B) 0.1 mg/l BAP, 0.01mg/l TDZ). The results indicated that the minimum callogenesis was observed in nodal and petiole explants in NT medium enriched with A hormonal combination. Callogenesis, fresh and dry weight of bark, stem and leave explants were significantly decreased in WPM and NT media containing B hormonal combination as compared with A hormonal combination. The maximum callogenesis was observed in bark explant in WPM (81.14) and NT (70.36) media with A hormonal combination as compared with other explants. The average fresh and dry weight of this explant in NT medium was more than that of WPM medium with the same hormonal combination.
K. Mahdavi Mashaki; A. Moieni; M. Jalali Javaran
Abstract
Rose is known as the queen of flowers. Rosa damask (Rosa damascena Mill.) is utilized as a main source of rose oil and rose water in Iran. Biotechnology methods are considered as a fast and efficient way to produce haploid plants and pure lines. Among the different methods, androgenesis is considered ...
Read More
Rose is known as the queen of flowers. Rosa damask (Rosa damascena Mill.) is utilized as a main source of rose oil and rose water in Iran. Biotechnology methods are considered as a fast and efficient way to produce haploid plants and pure lines. Among the different methods, androgenesis is considered as the most effective procedure, due to its high quantity of microspores in the anthers. In the present study, the effect of medium composition was evaluated on anther culture in two ecotypes of Damask rose, and miniature rose. The results showed that the interaction between H1 medium and mid-uninucleate stage produced the highest callogenesis in Damask rose, Kashan ecotype. Furthermore, the results showed that removing ammonium nitrate and doubling potassium nitrate in the medium produced higher callogenesis. In miniature rose, the medium containing calcium chloride and calcium nitrate produced higher callogenesis. Also, the amino acids improved callogenesis in anther culture of Damask rose, and miniature rose. Moreover, glycine, glutamine and casein hydrolysate were more effective than other studied amino acids. Sucrose was a better sugar compared to sorbitol for callogenesis in the studied genotypes. The chromosome counting and flowcytometery results illustrated that the produced calli were tetraploid (28 chromosomes).
P. Sarkheil; M. Omidi; S.A. Peyghambari; S. Davazdahemami
Abstract
Seeds were cultured on Whatman paper by sterile water in cube, solid MS and solid MS. Seeds were not germinated on medium but 80% of seeds were germinated on Whatman paper, so this method is used as a basic method. Seeds were germinated after four days and after two weeks of culture they had normal roots, ...
Read More
Seeds were cultured on Whatman paper by sterile water in cube, solid MS and solid MS. Seeds were not germinated on medium but 80% of seeds were germinated on Whatman paper, so this method is used as a basic method. Seeds were germinated after four days and after two weeks of culture they had normal roots, shoots and leaves. Cultures were incubated at 25◦ ±2◦C and exposed to 16 hours light per day. Explants were cultured on MS medium supplemented with 3% sucrose and solidified with 0.8% (w/v) agar. The pH of the medium was adjusted to 5.8 before autoclaving. Different types of explants were used for this experiment; root, crown, apical meristem, hypocotyls and leaf. Between different kinds of explants leaf didn’t response to callogenesis. The effects of different combinations of 2,4-D (2,4-Diclorophenoxy acetic acid) and BAP (6-Benzylaminopurine) were studied. Subculture was done every 3 weeks. In order to determine regeneration ability, the initiated callus were transferred to a regeneration medium which was composed of macronutrient, micronutrient and organic components of MS, 2,4-D (0.2 mgL-1), BAP (0.5, 2, 4, 15, 20 and 25 mgL-1) and MS without hormones, 0/3% sucrose, pH 5.8 for 4 weeks. In the presence of 2,4-D (2 and 4 mgL-1) and BAP (0.25 and 0.5 mgL-1) in the callus induction medium, high callus production percentage was reported. The hypocotyls, in contrast to the primary leaf explants, and apical meristem segments were more responsive to the tested combinations of 2,4-D and BAP. The callus from all explants was soft, watery and loose friable. During subculture period, hypocotyls and apical meristem were proliferated more on medium with the addition of (0.25 and 0.5 mgL-1) BAP and (2 and 4 mgL-1) 2,4-D than the medium contain BAP (1 mgL-1) and 2,4-D (8 mgL-1). The present study, in F. vulgare MS media without any hormone was sufficient to regenerate the plantlet from the hypocotyls, roots and apical meristems explants. In MS medium supplemented with BAP (0.5, 2 and 4 mgL-1) and 2,4-D (0.2 mgL-1) shoots were formed earlier when the number of subculture was increased 4 times.
G. Ghazian Tafrishi; M. Azizi; M. Farsi
Volume 22, Issue 3 , November 2006, , Pages 172-179
Abstract
St Johns Wort (Hypericum perforatum L.) from Hypericaceae is an important medicinal plant, which its secondary metabolites, hypericin and hyperforine, have several medicinal effects such as antidepressant, antiviral, antibacterial and etc. The importance of studying in vitro culture of medicinal plants ...
Read More
St Johns Wort (Hypericum perforatum L.) from Hypericaceae is an important medicinal plant, which its secondary metabolites, hypericin and hyperforine, have several medicinal effects such as antidepressant, antiviral, antibacterial and etc. The importance of studying in vitro culture of medicinal plants is optimizing these protocols for subsequent studies about effective factors on biosynthesis of secondary metabolites and applying these methods in improving medicinal plants. Since now there are no report on in vitro culture of Iranian St Johns Wort and for the first time we studied the callogenesis, shoot regeneration and rooting process of this plant. The seeds of Iranian St Johns Wort were collected from Ardebil province and the base growth media was MS and for callogenesis we studied the effect of several levels of 2,4-D (0.25, 0.5, 1, 2 and 5 mg/l) and BA (0.25, 0.5, 1 mg/l) or KIN (0.25, 0.5, 1, 2 and 5 mg/l) and NAA (0.5, 1, 2 and 5 mg/l). Several levels of BA (0.25, 0.5, 1and 5 mg/l) and several levels of KIN (1, 2 and 5 mg/l) accompanied by 1 mg/l NAA were used for shoot regeneration in callus. Several levels of NAA (0, 0.5, 1, 2 and 5 mg/l) were used for rooting of the shoots. The growth condition was 25°c and 16/8 hours period for rooting and shoot regeneration, darkness for callogenesis. The results of callogenesis with Duncans Multiple Range Test at 5% showed that highest callus fresh weight (2.1937 gr) was obtained in 0.25 mg/l 2,4-D with 1 mg/l KIN. Results of shoot regeneration in level 5% showed that maximum number of shoots (95 shoots/call and 4 Cm length) obtained in treatment contain 1 mg/l NAA with 1 mg/l KIN. Results also showed that maximum root number (4.7 roots per shoot and 2.2 Cm length) was in hormone free media.
