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Leaf and root explants from 10 to 20 day– old seedlings of Plantago lanceolata, were cultured on MS, MSH, NT and B5 basal media with various concentrations of different plant growth regulators. The best callus induction was obtained with 0.4 mgl-1 2,4-D in MSH I with 400 callus index. Callus growth was significantly stimulated (P=0.05)with 0.8 mgl-1 2,4-D and 0.1 mgl-1 Kin in MSH II, with 799 mg fresh weight and 79 mg dry weight. A higher production of mucilage was obtained in MSH II. The mucilage content of different media varied from 10.40 to 14.73% dry weight. Perusal of the data reveals significant differences with regard to the total mucilage content. Callus has nearly 3 times and 1.5 times more mucilage than seeds and leaf and root parts respectively.

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Water stress is a serious environmental problem throughout the world. It is a conse-quence of both drought and salinity which may be relieved by breeding cultivars that can tolerate low soil water potentials. Alfalfa (Medicago sativa L.) is normally grown in arid and semi-arid zones. One approach to improving the ability of the crop tolerate drought is to make use of the variation provided by tissue culture derived from somaclonal varia-tion. Seedlings of a moderately salt tolerant line CUF 101-1S derived from the commercial variety CUF 101 were used as a source of explants to initiate and regenerate tissue cul-tures. Regenerant plantlets were allowed to grow to maturity, selfed, and set seed. These seeds (R1) were germinated and grown in a nutrient medium containing 0, 200 and 250 g l-1 polyethylene glycol (PEG) 6000 and additional calcium as CaCl2 (4.0 mol m-3) for 14 days to assess their ability to withstand stress in the progeny generation. Tolerance to PEG was assessed using measurements of root and shoot lengths. Proline levels and activ-ity of the antioxidant enzymes, catalase (EC 1.11.1.6) and glutathione reductase (GR) (EC 1.6.4.2) were also determined. Selected somaclones were more tolerant than the parent. A large increase in the level of proline was observed in the somaclones compared with the parent variety in response to PEG stress. The activity of catalase and glutathione reduc-tase increased in the tolerant genotypes but remained unchanged in the parent when they were subjected to PEG stress.

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Parsi Zira, Bunium persicum (Boiss.) B. Fedtsch., or Black Zira, a member of the Umbellifera family, is one of the most important medicinal species with high economic value. Generally, there is limited information about in vitro culture of Bunium persicum. In this study, the effect of various media (MS, B5) and combinations of plant growth regulators, NAA and 2,4-D alone or together with Kin, on callus induction and shoot regeneration from hypocotyl explant were investigated. This experiment has been carried out in a completely randomized design with 30 treatments and 10 replications per treatment. The results showed significant effects of treatments on regeneration and callus induction. All the measured traits for MS medium were superior. The highest callus frequency was observed on the medium containing 0.1 mg L-1 or 1 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D) as well as 2 mg L-1 2,4-D and 0.5 mg L-1 kinetin. The best treatment for somatic embryogenesis was the medium containing 2 mg L-1 2,4-D. The best response for shoot regeneration was observed on the medium supplemented with 1 mg L-1 2,4-D. The medium containing 1 mg L-1 2,4-D was the best for callus induction and shoot regeneration simultaneously. The regenerated shoots were rooted on basal medium.

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The use of healthy planting material is an important prerequisite for controlling destructive viral diseases in potato production. Virus elimination methods such as meristem culture and thermotherapy are usually used to produce the nucleus stock of healthy planting material. Here, we report a new technique of electrotherapy for elimination of two potyviruses, Potato virus A (PVA), and Potato virus Y (PVY), from potato plants. Electric currents of 15, 25 and 35 mili Ampers for 10 to 20 minutes were used for eliminating the combination of PVA and PVY in six potato cultivars. An electric current of 35 mili Ampers for 20 minutes was the most effective electrotherapy treatment for eliminating these two viruses. Responses of potato genotypes to electrotherapy were significantly different. Regenerations of electrotherapy treated plantlets in cultivars Lady Roseta and Banaba were 70.8 and 66.6 percent respectively, while these were approximately 54 to 58 percent for cultivars Olimpia, Agria, Desirea and Clone 69. Virus free plantlets were successfully produced in the potato cultivar Lady Roseta. However, in other cultivars examined in this study, electrotherapy resulted in drastic reduction of virus concentration but not total eradication of the virus infection from plant tissues.

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Nowadays, the use of tissue culture technique in the production of plant secondary metabolites, such as phenolic compounds, as a natural food additive has become very widespread in the food industry. The aim of this study was to investigate and compare the content of phenolic compounds and antioxidant activity of grapevine Rasha and Qzel Ozum cultivars that were produced by tissue culture under in vitro condition. For this purpose, the terminal buds of grape cultivars were cultured in Murashige - Skoog Medium to produce complete seedlings. Then the methanolic extract was prepared from healthy grapevine leaves and to perform polyphenol composition tests using high performance liquid chromatography with ultraviolet detector, functional groups using Fourier-transform infrared spectroscopy, total phenol content, total flavonoids, total anthocyanins, total carotenoids, acid content, ascorbic acid, free radical scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH), and hydroxyl  (OH⁻). The results showed that the amount of total phenolic compounds, total flavonoids, total anthocyanin and total carotenoids of methanolic extracts of Rasha cultivar were 50.35±0.10, 29.72±0.05, 10.41±0.20, and 1.82±0.50 mg/g, respectively, and in Qzel Ozum cultivar were 23.06±0.31, 20.76±0.15, 7.28±0.15, and 94.94±0.42 mg/g, respectively. Ascorbic acid, DPPH and OH⁻ free radicals scavenging activity in methanolic extracts of Rasha leaves were 1.83±0.04, 25.38±1.52, and 75.04±2.74 %, respectively, and in Qzel Ozum leaves were 0.85±0.01, 18.71±2.05 and 68.96±3.61 %, respectively. As a general result, the results showed that the grape leaves of the Rasha cultivar were more than the grapevine leaves of Qzel Ozum cultivar in terms of all studied characteristics (p <0.05). This study also showed that grape leaves are rich in phenolic compounds and natural antioxidants and have the potential to be used in the food and pharmaceutical industries.

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Saffron (Crocus sativus L.) is a sterile species and biotechnological method is suggested to improve different characteristic in this valuable plant. In this study, an efficient protocol was provided for callus induction and regeneration of saffron using thin cell layer explants. Longitudinally and transversally, thin cell layer explants with approximately 1 mm thickness of apical buds were cultured on MS medium supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D), 6-BenzylAminoPurine (BAP) and 1-NaphthaleneAcetic Acid (NAA). The highest amount of callus induction (100%) was obtained from transverse thin cell layer explants of apical bud in MS medium containing 2 mg L^-1 BAP and 2 mg L^-1 NAA during 3 months incubation under dark condition at 20°C. The maximum percent of shoot regeneration (75%) was observed on the MS medium containing 0.5 mg L^-1 BAP. The results of this investigation revealed that the thin cell layers from buds are suitable explants for regeneration.

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Using immature embryos that undergo somatic embryogenesis, we studied the effects of different hormonal compounds and media on controlling secondary somatic embryogenesis (SSE) in Persian oak (Quercus brantii L.). To this end, we focused on the immature embryos that were subjected to several treatments including chilling (at 4 ℃) period and SH, MS, 2,4-D, IBA, BAP, and glutamine concentrations in 5 separate sequential experiments. The results showed that, by extending chilling period to 8-weeks, SSE induction was reduced (68.75%). In different MS concentrations or MS containing PGRs, the lowest globular embryo (66%) and the secondary embryo induction (87.5%) were observed in embryos treated with MS+IBA+BA. Adding 0.75 mg L^-1 glutamine to MS resulted in a decrease in the secondary somatic embryogenesis (56.25%). Among MS and SH media, 1/2 SH almost entirely controlled this phenomenon (6.25%). The highest maturation progression was obtained in the SH+glutamine treatment, which had the highest conversion to plantlet percentage (100%) and vigor index of plantlets (51.93) compared to the use of SH alone. We found that nutrient and PGR concentration were critical in embryo maturation and conversion percentage and stop the embryo induction cycle that plays a major role in secondary embryogenesis.
 

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Digitoxin is a glycoside isolated from leaves of various species of Digitalis genus that are commonly called foxgloves. Digitoxin is used for chronic cardiac insufficiency, tachyarrhythmia form of atrial fibrillation, paroxysmal ciliary arrhythmia, and paroxysmal supraventricular tachycardia. 1-Deoxy-d-Xylulose 5-phosphate Reductoisomerase (DXR) is an important rate-limiting enzyme in 2-C-Methyl-d-Erythritol 4-Phosphate (MEP) pathway, and expression level of DXR plays a critical role in control of plant isoprenoid biosynthesis pathway. The effects of different elicitors including putrescine, spermine, and spermidine at concentrations of 50 and 100 mg L^-1 and also Methyl Jasmonate (MJ) at 50, 100, and 200 μM concentrations were explored on transcript levels of DXR gene in cell suspension culture of foxglove (Digitalis purpurea). DXR transcription levels were assessed by the semi-quantitative RT-PCR. Results showed that elicitors had significant effects on the expression level of DXR and contents of cardenolide and digitoxin. The highest digitoxin (61.3 μg g^-1 DW) and cardenolide (1.48% mg^-1 DW) content was observed in 100 mg L^-1 spermidine treatment. In all treatments, the expression of DXR gene was increased compared with the control sample. The highest transcription levels of DXR gene was observed in 50 μM MJ and 100 mg L^-1 putrescine elicitors, which was nine folds higher than the control condition, and the lowest levels were observed for putrescine treatments at concentrations of 50 and 100 mg L^-1, respectively.