ولی الله قاسمی عمران

Gotu Kola (Centella asiatica L.) has been listed in threatened species in Iran. It has been used for treatment of a wide number of health disorders. The herb was used for wound healing, anticarcinogenic, mental illness and revitalizing the nerves and brain cells. In vitro culture and micropropagation are basic tools for conserving this medicinal plant. The purpose of this study is to achieve a suitable protocol for propagation of this plant in in vitro. In the first experiment, interaction of different concentrations of BAP (0, 0.5, 1, 1.5, 2.5, 3.5 mgl-1) and NAA (0, 0.25, 0.5, 1, 2 mgl-1) were examined on leaf explants. The results of the callogenesis experiment showed that MS medium with 1.5 mgl-1 BAP and 0.5 mgl-1 NAA, produced the best callus. In the next step, the interaction of BAP at 0, 1, 2, 3 mgl-1 and IBA at 0, 0.5, 1 mgl-1 was investigated in proliferation of node explants. Rooting of amplified explants was investigated by using two types of auxin IBA and NAA. The results of the micropropagation experiment indicated that MS medium of 2 mgl-1 BAP and 1 mgl-1 IBA was determined the most suitable medium for micropropagation. In the comparison of the effect of auxin treatments in MS medium on rooting, MS medium supplemented with IBA 1 and 2 mgl-1 were chosen as the best rooting treatment.

Light-emitting diodes (LEDs) can be used as a useful alternative to in vitro culture of various plant species, which not only accelerates plantlet growth and development but also reduces illumination costs. Here, the effects of different light treatments (fluorescent (control), blue LED, red LED, 70% blue/30% red, 70% red/30% blue and SMD lights) on protocorm-like bodies (PLB) growth and development, photosynthetic pigments, antioxidant enzyme activity, and the content of protein and soluble sugars of Phalaenopsis pulcherrima obtained from protocorm explants on ½ MS (Murashige and Skoog) or VW (Vacin and Went) with or without growth regulators (IBA+BAP) were evaluated. The results showed that LED and SMD treatments improved the induction and growth of PLBs compared to fluorescent treatments, which had the highest numbers of PLBs and biomass under blue and red LED light, respectively. Under the same light treatments, the highest numbers of PLBs and biomass were obtained on ½ MS+IBA+BAP. Compared to fluorescent light, LED and SMD lights increased the content of photosynthetic pigments. Among the culture media used, the highest content of photosynthetic pigments was observed on ½MS+IBA+BAP. Compared with fluorescent treatment, a significant increase in the activity of antioxidant enzymes and the content of protein and soluble sugars was observed under LED and SMD treatments, with the largest increase recorded under blue LED. Therefore, our results proved that the use of LEDs can be a practical and effective way to increase the induction of PLBs and improve photosynthetic pigments in the in vitro culture of P. pulcherrima.

Luffa (Luffa cylindrica) is a plant from the Cucurbitaceae family that grows mostly in tropical and subtropical regions, as well as in most regions of Iran. In this research, the genetic diversity of nine native and non-native genotypes of L. cylindrica was investigated through the evaluation of the chloroplast trnH-psbA intergenic region (IGS). After sampling the young leaves, DNA extraction was performed by using the Dellaporta method, and PCR was conducted by using IGS intergenic region primers. After sequencing of the amplified products, their quality was determined using Chromas software and then aligned using ClustalW method by BioEdit and MEGA7 softwares. Next, the dendrogram of phylogenic relationships was drawn and the matrix of the difference and similarity of the sequences were determined. In the present research, by analyzing the relationships between studied samples, based on the trnH-psbA (IGS) marker, a strong intraspecies variation was observed in native and non-native L. cylindrical genotypes. The genetic distance matrix between the samples examined in this research ranged from 0 to 6.865 with an overall average distance of 2.53. The average value of synonymous and non-synonymous substitutions (dN/dS) for the IGS sequence was ds/dn = 0.68, which indicates positive and pure line selections in the process of natural selection of studied genotypes. The results of this research showed that trnH-psbA is a suitable marker for evaluation of the intraspecific diversity of luffa species.

Centella asiatica, is well known to be a valuable medicinal plant for producing valuable compounds such as asiaticoside, asiatic acid, madcasoside and madcasic acid. The plant is believed to improve memory, lower blood pressure, be a strong antioxidant and anticancer. Therefore, it is important to optimize tissue culture methods in order to facilitate the extraction of medicinal compounds, gene transfer as well as improvement of medicinal properties of the plant. Calli prepared from various medicinal plants can be used to increase the amount of medicinal compounds in the cell suspension culture and gene transfer. The aim of this study was to investigate the effect of different concentrations of two hormones, BAP and NAA, on leaf explant for callus initiation, as one of the important sources of secondary metabolites production. For this purpose, leaf explants were treated with 6 different concentrations of BAP (0, 0.5, 1, 1.5, 2.5, 3.5 mg/L) and 5 different concentrations of NAA (0, 0.25, 0.5, 1, 2 mg/L). The results of this study showed that the best callus was obtained by the combination 1.5 mg/L of BAP and 0.5 mg/L of NAA, resulting in the leaf explants with callus induction of 100%, fresh weight of 1.457 gr and callus diameter of 1.459 cm. The results showed that two hormones of BAP and NAA have synergistic effects on the increase of the quality and quantity of the produced calli.

Centella asiatica (L.) Urban, a medicinal plant from Apiaceae family, is known as an endangered species in Iran. This plant has many therapeutic properties, including memory enhancement, anticancer, and treating skin diseases and has also high antioxidant activities. Plant cell and tissue culture is an important tool in basic studies and has commercial applications. Tissue culture method can be used to breed medicinal plants or change the amount of secondary metabolites. Due to the high value of C. asiatica, in vitro cultivation can be used to produce more of this plant. Optimizing tissue culture is useful for conducting the applied researches and metabolic engineering of different secondary metabolites biosynthetic pathways. The present study was aimed at achieving a suitable protocol for in vitro propagation and determining the appropriate concentrations of growth regulators for micropropagation of this valuable species. The interaction of BAP (0, 1, 2, and 3 mg.l-1) and IBA (0, 0.5, and 1 mg.l-1) was investigated on proliferation of nodal explants in MS medium. Then, rooting of the propagated explants was evaluated using IBA and NAA. The results showed that MS with 2 mg.l-1 BAP and 1 mg.l-1 IBA and MS with 1 mg.l-1 IBA were the best media for proliferation and rooting, respectively.

Luffa is a plant from the Cucurbitaceae family with the scientific name of Luffa cylindrica. In addition to the food consumption of mature green fruit and the medicinal and health properties of seed oil, it is important due to the fibrous spongy texture of its dried fruits. There is a great variety in terms of shape, size, quality of sponge, etc. among different cultivars of Luffa, the study of which helps plant breeders to identify the genetic capacity of traits associated with its important breeding goals.

In this study, the genetic diversity of 5 indigenous Lufa genotypes related to Mazandaran, Kerman, Sistan and Baluchestan provinces, and 3 non-Indigenous genotypes related to China, Brazil and Afghanistan were studied by evaluating the ITS (Internal transcribed spacer) ribosome region. After sampling of young leaves, DNA was extracted by Dellaporta method, and PCR was performed using a pair of ITS primers. After sequencing and qualification, amplified products were aligned and dendrogram of phylogenic relationships and difference and similarity matrix of sequences were determined.

In this research, from a total of 703 sites, 175 sites had deletion and addition (170 monomorphes and 5 polymorphs) and 528 sites were lake of deletion and addition. Most of the substitutes for the ITS sequence were transversion (55.5%), and only 44.5% of the substitutes were transitional type. Also, the mean ratio of thymine, cytosine, adenine and guanine was 9.18, 8.28, 19.9 and 42.4% of the total nucleotides. The average numerical value of DN / DS ratio was 0.86. The overall average of the genetic distance between the samples examined, for the ITS marker, using the Maximum Composite model was 0.107.

Studying the relationships between tested genotypes based on the ITS region, as analyzed in this study, showed the presence of intra-species variation in the genotypes of Luffa aegyptiaca studied. However, relatively low genetic distances between genotypes may indicate the relative weakness of this marker in the genetic studies of Luffa. Therefore, for a more accurate conclusion it is suggested that this marker is be used as a supplementary barcode with other markers.

Genetic relationships among twenty-three Chickpea genotypes include seven Chickpea cultivars, and 16 wild genotypes were analysed by 10 ISSR markers. On average, 11 bands per primer were observed, and 83.4 % polymorphic bands were shared in ISSR analysis. The results of clustering demonstrated the efficiency of ISSR markers (particularly ISSR1 (UBC-834) and ISSR8 (UBC-864)) for reporting appropriate parents for breeding programs to biotic and abiotic stresses. Principle Coordinate Analysis (PCoA) clustered 23 Chickpea genotypes in two groups by which six cultivars separated in one group. Genetic data extracted from ISSR analysis are practical in Chickpea to choose parental sources for inbreeding programs.

Chilling injury (CI) is one of the main factors involved in reducing pomegranate quality during cold storage. In the present study, the interaction between 1-MCP and three different coatings (cellophane, confectionery waxy paper and 1%, 2% sodium alginate) on maintenance of postharvest quality of pomegranate fruits were investigated during 120 days of cold storage .The results showed that uncoated fruits had higher weight loss and total soluble solids (TSS) than coated fruits during storage. The results also showed that untreated (control) fruits showed higher weight loss, decay and total soluble solids (TSS) than 1-MCP treated fruits during cold storage. Cellophane had a significant effect on delaying ion leakage and preventing weight loss of pomegranate fruit.The 1-MCP+cellophane and 1-MCP+confectionery wax paper could reduce chilling injury by delaying the reduction of bioactive compounds such as total phenolics and antioxidant activity. The lowest decay severity was also observed in fruits treated with 1-MCP. In general, it can be concluded that postharvest application of 1-MCP+cellophane was the most effective treatment to induce chilling tolerance and extend the storage life of pomegranate.

The production of ornamental cultivars with new colors is the main goal in the flower and ornamental plants industry. African violet is well known commercially and available in different colors except yellow flower. The aim of this study was to change the color of A. violet petals and to produce the biosynthetic pathway of auron pigment through genetic manipulation of AS1 and 4'CGT genes. To this end, t genes were isolated from Antirrhinum majus petals yellow using PCR with specific primers and ligated into the expression vectors pCAMBIA1304 and pBI121, respectively. Recombinant pBI121+4'CGT and pCAMBIA1304+AS1 vectors were confirmed using colony PCR, enzyme digestion, sequencing and comparative analysis in the database. They were then transferred to Agrobacterium strain LBA4404 by electroporation method and was examined in the petals of A. violet using a temporary expression system. In this system, Agrobacterium suspension carrying the gene structure was injected using a syringe at the base of the petals.. Morphological results after 3 days clearly showed discoloration of white to pale yellow petals and no change was observed in control petals. PCR analysis and light microscopy confirmed gene expression and phenotypic changes, respectively. Also, aureusidin-6-O-glucoside (AOG) was detected in transgenic petals using HPLC-DAD-MSn. By using this system, the gene structure can be transferred to ornamental plants with more confidence for permanent transgenic. In addition, this research can open the way for engineering the creation of yellow color in ornamental plant species that do not have this type of color.

In recent years, the use of natural and healthy compounds has been considered as a new method to control chilling and maintain postharvest quality of horticultural products. In this study, for the first time, pomegranate fruits were immersed in L-arginine solution at concentrations of 0, 1 and 2 mM and Its effects on the quality of pomegranate fruits ‘Malas-e-Saveh’ grown in Sari region was evaluated during 120 days in cold storage. Based on the obtained results, the treatment significantly increased the total phenol and antioxidant properties of the fruit compared to the control. Fruits treated with 1 mM L-arginine showed more antioxidant activity compared to the control. Moreover, the activity of antioxidant enzymes (CAT, SOD, APX) as well as PAL enzyme increased accordingly. Whereas, H2O2 accumulation and PPO enzyme activity in fruits treated with 1 mM L-arginine were significantly reduced. Based on our results, L-arginine treatment can be used as a useful and practical method to maintain nutritional quality and increase the pomegranate storability due to its safety and effectiveness.

The mutation in the meristem layers creates different genetic backgrounds (chimera) in the plant tissue. The mutation in L1 layer of shoot apical meristem generates a periclinal chimera. UF3GT is an effective enzyme in floral coloration, inducing anthocyanin accumulation in petals. This study investigates direct and indirect regeneration systems and different explants to propagate two cultivars of periclinal chimera (Saintpaulia ionantha), namely Taro taraneh and Aghaz, using in vitro culture. The evaluation of UF3GT gene expression pattern by Real-Time PCR revealed the role of anthocyanin accumulation in the petal coloration of chimera plants. Results pertaining to both cultivars showed that inflorescence and leaf explant had the highest and lowest percentage of pinwheel phenotype, respectively. In addition, mutant characteristics were faded in the leaf regeneration of periclinal chimera. Furthermore, the highest percentage of periclinal chimera was generated in direct regeneration. Gene expression analysis revealed that UF3GT was expressed in the colorful part of chimera petal, while UF3GT expression was significantly reduced in the muted part. HPLC chromatogram also detected that cyanidin and delphinidin components were not present in the white part of either cultivar. The anthocyanin biosynthesis pathway appears to be blocked and anthocyanin accumulation does not occur in the petals. Inflorescence is likely to induce a pinwheel pattern in regenerated plants, probably owing to its lateral bud. It seems that different meristem layers are associated with the formation of epidermis and induce pinwheel phenotype.

African violet is a commercial ornamental plant with various colors. For direct regeneration, in the first experiment, the effect of leaf, node, petiole, root, petal and sepal explants of four African violet cultivars were evaluated on MS medium containing different concentrations of NAA+BAP and IBA+BAP hormones, which was performed as a factorial in a completely randomized design. In the second experiment, the tolerance threshold of leaf and petiole explants to Hygromycin and Kanamycin antibiotics was evaluated by culturing on selective media supplemented with different concentrations of given antibiotics. After determining the suitable concentrations of the antibiotics for selective medium, the transformation of AS1 and 4'CGT genes was conducted with Agrobacterium LBA4404 strain containing pCAMBIA1304 and pBI121 plasmids, respectively. PCR method was used for the detection of transgenic plants. First experiment revealed that the leaf explant was the best sample and MS medium containing 1 mg L-1 IBA+1 mg L-1 BAP was the best combination for in vitro regeneration of all genotypes. The results of the second experiment showed that the M4 medium (containing 50 and 75 mg L-1 Kanamycin and Hygromycin, respectively) completely inhibited the regeneration of non-transformed samples. Finally, transgenic plants were selected using PCR with specific primers for transferred genes.

In this study, the genetic diversity of nine different indigenous and non-indigenous luffa cultivars with the scientific name Luffa cylindrical was evaluated by analyzing the sequence of chloroplast rbcL gene. After DNA extraction and PCR using rbcL gene-specific primers, the amplified products were sequenced and aligned, and the dendrograms of phylogenetic relationships were plotted. Of the 703 sites identified in the study, 179 sites had deletions and additions (177 monomorphs and 2 polymorphs) and 524 sites without deletions and additions. Also, the average numerical value of the dN/dS ratio was approximately one. Overall, the results of this study showed that according to the dendrogram obtained from cluster analysis, and genetic distances, not only with the help of rbcL marker, the diversity within species of this plant species can be well evaluated, but these genotypes can be to some extent classified based on geographical area. It is suggested that the diversity of these genotypes be examined by other DNA barcodes and compared with the results of the present study.

Nowadays, biotechnological methods such as transgenic hairy root culture and application of elicitors have become an attractive source for secondary metabolites production. Hyssop species (Hyssopus spp.) having active ingredients including antioxidants such as phenolics, in this plant have shown anti-mutagenic, anti-carcinogenic and antiglycemic effects.

The present study was conducted to investigate the effect of silver nanoparticles on the growth and antioxidants of transgenic hairy root in two Hyssopus species including H. officinalis and H. angustifolius.

In this experiment, we used silver nanoparticles concentrations of 0, 0.1, 0.2, 0.3 mg/l on transgenic hairy roots and Agrobacterium rhizogenes ATCC15834 strain for induction of hairy roots. Extraction of the samples were performed following ultrasonic method. Total phenolics content were determined using spectrophotometry. The antioxidant activity (AOA) of the extract were evaluated through 2, 2-dipheny l-1-picryl-hydrazy l (DPPH) method.

The results showed that silver nanoparticles as an elicitor had significant effect on the measured traits. The highest dry weight and fresh weight of the transgenic hairy roots were observed as much as 3.8 and 16 g/l in H. angustifolius and 3.2 and 14 g/l in H. officinalis, respectively. Which were caused by silver nanoparticles at the concentration of 0.1 mg/l, The amount of total antioxidants in H. angustifolius and H. officinaliswere 90.81% and 89.84%, respectively. In this study, we could observe relationship between antioxidant activity and plant phenolic content.

In general, we found that 0.1 mg/l silver nanoparticles could improve the growth of transgenic hairy roots and increase their phenolics and antioxidants content.

In the present study, hairy root induction from leaf and stem explants in two Hyssopus species (H. officinalis and H. angustifilius) was assessed using four Agrobacterium rhizogenes strains including A4, ATCC15834, LB9404 and 2656 on three culture media (MS, ½ MS, B5) as a factorial experimental based on a completely randomized design with three replicates. Almost all leaf and stem explants produced hairy roots. A fragment of transferred rol B was amplified through PCR from the genomic DNA extracted from transformed roots. Different A. rhizogenes strains and culture media and Hyssopus species had significant effects on hairy root percentage and root length. The highest rooting percentage (%71) and root length (4.7 cm) were resulted from stem explants of H. angustifolius co-cultivated with the strain ATCC15834 on MS culture medium. While, the leaf explantsof H. officinalis co-cultivated with the strain A4 on MS culture medium showed the rooting percentage of 55% and the root length of 1.6 cm at most. The MS medium was the best culture medium for both species. These results can be useful in genetic manipulation of hyssop and its hairy root culture to produce high-value secondary metabolites.

In order to investigate the role of Piriformospora indica endophyte fungi on some vegetative, physiological and morphological parameters along with antioxidant activity of stevia (Stevia rebaudiana) under cadmium (Cd) stress, an in vitro experiment was conducted. Experiment arranged in completely randomized design with three replicates. Treatments were five levels of Cd (0, 5, 10, 15 and 20 mg/L from cadmium chloride source) and two levels of fungi inoculation (including control and inoculation with P. indica). After 30 days, some growth characteristics, photosynthetic pigments, antioxidant enzymes activity, electrolyte leakage and soluble protein were measured. Regression analysis indicated that among vegetative dry weights, stem dry weight in uninoculated plants showed the maximum sensitivity (76%) to Cd concentration gradients. Fungi inoculation, however, markedly improved root dry weight up to 57%. Among morphological parameters, leaf area showed the highest reduction when Cd increased in the growing medium. By contrast, where P. indica was inoculated, green leaf percentage (51% reduction) was the most tolerant parameter. In terms of photosynthetic pigments, fungi inoculation could ameliorate reduction of carotenoid and chlorophyll a/b contents from 77 and 37 to 40 and 11%, respectively. In conclusion, results of the present study indicated that P. indica inoculation particularly at lower Cd concentrations could relatively improve stevia tolerance throughout decreasing H2O2 accumulation and enhancing photosynthetic pigments.

In order to evaluate the effect of Piriformospora indica fungi inoculation on antioxidant systems and photosynthetic pigments of stevia medicinal plant under salt and drought stresses, an in vitro experiment was conducted. Experiment arranged in factorial based on completely randomized design with three replicates. Treatments were three levels of osmotic potential (0, -5, and -10 bar), three osmotic sources including NaCl (Na), Mannitol (M) and NaCl+Mannitol (N+M) and two levels of fungi inoculation (uninoculated control and inoculation with P. indica). Results showed that among the mentioned osmotic sources, NaCl had the most inhibitory effect on photosynthetic pigments of stevia. When M concentration increased in uninoculated plants, the enzymes activitiy such as catalase and superoxide dismotase increased too. In non-stress conditions P. indica improved photosynthetic pigments including chlorophyll a, b, a+b and carotenoids. Also, inoculation of stevia plantlets with P. indica at osmotic level of -5 bar caused by M+Na markedly improved chlorophyll b and a+b up to 41 and 57%, respectively as compared to the uninoculated control. Fungi inoculation also improved activity of antioxidant enzymes as compared to uninoculated control. Where Na and M+Na was applied to cause -5 bar of osmotic level,  superoxide dismutase activity increased by 62 and 185%, respectively as compared to the control. In conclusion, results showed a positive effect of P. indica on stevia tolerance to both salinity and drought stresses.

The present study was performed to evaluate the effects of different concentrations of ethyl methane sulfonate (0, 0.1, 0.2 and 0.5%) on some physiological characteristics of regenerated plants from calli of stevia at 30, 60 and 120 min under various levels of salinity stress (0, 50 and 100 mM of NaCl). This experiment was carried out based on completely randomized two-factorial designs with three replications. With respect to the result, the regenerated calli became dark and hidden in the medium under exposure time of 120 min, the length of stem regenerated calli was increased under exposure times of 30 and 60 min. Moreover, our data showed that EMS mutagenesis had a significant effect on physiological traits of regenerated stevia under salinity stress at the probability level of 1%. Consequently, the stevia mutants of M10, M11, and M19 showed the highest resistance to different levels of salinity which can be considered as potential samples for further breeding programs.

In order to evaluate the effect of Piriformospora indica fungi inoculation on antioxidant systems and photosynthetic pigments of stevia medicinal plant under salt and drought stresses, an in vitro experiment was conducted. Experiment arranged in factorial based on completely randomized design with three replicates. Treatments were three levels of osmotic potential (0, -5, and -10 bar), three osmotic sources including NaCl (Na), Mannitol (M) and NaCl+Mannitol (N+M) and two levels of fungi inoculation (uninoculated control and inoculation with P. indica). Results showed that among the mentioned osmotic sources, NaCl had the most inhibitory effect on photosynthetic pigments of stevia. When M concentration increased in uninoculated plants, the enzymes activitiy such as catalase and superoxide dismotase increased too. In non-stress conditions P. indica improved photosynthetic pigments including chlorophyll a, b, a+b and carotenoids. Also, inoculation of stevia plantlets with P. indica at osmotic level of -5 bar caused by M+Na markedly improved chlorophyll b and a+b up to 41 and 57%, respectively as compared to the uninoculated control. Fungi inoculation also improved activity of antioxidant enzymes as compared to uninoculated control. Where Na and M+Na was applied to cause -5 bar of osmotic level,  superoxide dismutase activity increased by 62 and 185%, respectively as compared to the control. In conclusion, results showed a positive effect of P. indica on stevia tolerance to both salinity and drought stresses. 

Drought is one of the most important abiotic agents that adversely affect the growth and development of plants. The arid and semi-arid regions where sesame (Sesamum indicum L.), as a relatively drought tolerant plant, is grown are specified by high temperatures. In order to evaluate the effects of paclubutrazul (PBZ) on the drought tolerant of sesame a factorial experiment were conducted based on randomized complete block design with three replications in 2014 at greenhouse of Educational Department of Kordkoy (Golestan province). Treatments included two genotypes of sesame (Yellow- white and Oltan), three levels of watering (20, 50 and 80% FC) and three concentrations of PBZ application (0, 7.5 and 15mg/L). Traits such as chlorophyll a, b and carotenoid contents, superoxide dismutase and peroxidase activity, yield and yield components were measured. Data were analyzed with SAS software. Results showed that genotype, water treatment and PBZ had a significant effect on chlorophyll a, b and carotenoid contents, superoxide dismutase and peroxidase activity of roots and leaves, yield and yield components. High PBZ application where plants exposed to the severe water stress significantly improved chlorophyll b (44%), superoxide dismutase of roots and peroxidase activity of roots and leaves. Yellow-white genotype showed higher chlorophyll a (30.4%), b (40.6%) and carotenoid concentration (29.9%), superoxide dismutase of roots (3times), superoxide dismutase of leaves (1.9 times) and peroxidase activity of roots (3times) in severe water deficit stress. Finally, it can be concluded that Yellow white responded better to drought stress than Oltan genotype with maximum levels of PBZ throughout antioxidant mechanismes.

Stevia rebaudiana is a medicinal plant belonging to Asteraceae family. This experiment was carried out to optimize the in vitro callus induction and plant regeneration of Stevia rebaudiana. The explants used in this study were leaf, shoot tip, nodal segments with two leaves and nodal segment without leaves derived from in vitro grown plantlets. The explants were transferred to PGR-free MS medium or supplemented with BA (0.1, 0.5 and 2mgl-1) solely or in combination with 0.5mgl-1 IBA and TDZ (0.1, 0.5 and 2mgl-1) solely or in combination with 0.5mgl-1 IBA. The highest callus percentage and callus volume induced with culturing leaf explant on MS medium containing 0.1 mgl-1 TDZ. The nodal segment without leaves cultured on MS medium containing 2mgl-1 BA + 0.5mgl-1 IBA produced the highest regenerated shoot number. The highest and lowest shoot length was observed in shoot tip cultured on MS medium containing 0.1mgl-1 BA + 0.5mgl-1 IBA and MS medium containing 2mgl-1 BA + 0.5mgl-1 IBA, respectively. Indirect shoot regeneration was observed only in leaf explant cultured on MS medium containing 0.5mgl-1 TDZ + 0.5mgl-1 IBA. The highest percentage of root induction was observed with culturing shoots on MS medium containing 0.1mgl-1 IBA.

To study intermittence of duration of exposure to magnetic field or MF (0.9 mT) on direct organogenesis of African violet explant in medium with and without PGRs these experiments performed. First experiment performed as completely randomized design (CRD) with three level of MF exposure (control, 5 day continuous and 5 day discontinuous) in PGR- free medium included via 6 repeat. The second experiment performed as factorial in CRD. Factors included auxin and cytokenin as two PGRs in 1mg Lit and 3mg Lit IBA and BAP levels and 3 levels of MF exposure as well as first experiment with 6 repeat. The results showed that explants exposed to a discontinuous MF with and without PGRs produced the highest shoot number (7.08) and the lowest number was produced in the control (2.35). Plantlets length was not affected by the MF but it was affected inside of PGRs. The leaf number in the continuous (4.4) and discontinuous (4.7) magnetic field was approximately doubled in comparison with control (2.77). The fresh and dry weight revealed better values in the MF discontinuous (0.49 and 0.03 mg) than continuous (0.35 and 0.02 mg) in PGRs-free medium. Results showed the fresh weight of plantlets was significantly higher for 1mgl-1 PGRs than 3 mgl-1 PGRs, when exposed to 0.9 mT MF for 5 days continuously, as well as for dry weight. It seems that the MF as a light abiotic stress can simulate the cell division and elongation and eventually the plant growth.

Hyssops are herbaceous and perennial plants, belonging to the Lamiaceae family. In this experiment, callus production of two hyssop species (H. officinalis and H. angustifolius called common and local, respectively) was investigated using two culture media and different levels of plant growth regulators (PGR’s) to achieve the best culture medium and PGR combination for producing callus as a starting plant material for plant regeneration and secondary metabolite production. To do this, the seeds were cultured in MS medium and after 2 weeks shoots were placed on to the MS and B5 medium and PGR treatments (control, 0.5mg/L 2,4-D, 1mg/L 2,4-D, 0.2 mg/L NAA + 0.5mg/L 2,4-D, 0.2 mg/L NAA + 1mg/L 2,4-D) and incubated in a controlled environment. Then, the callus induction time and percentage of callus producing explants, fresh and dry weight of callus and callus color were recorded. The results showed that interaction between species, medium and PGR treatments was significant at 1% level. Meanwhile, mean comparison analyses showed that the highest amount of callus production in both evaluated hyssop species was observed in MS media supplemented with 0.5 mg/L 2,4-D and 0.2 mg/L NAA, in which the common hyssop showed calli after 6 days, while the local hyssop species started callusing after 11 days. According to this research, this PGR treatment is proposed for callus induction in both species. Given the importance of these medicinal speciesof hyssop especially the local one and probability of its extinction, these results can be used for protecting this valuable herb.