کشت درون شیشه ای

 Lily, a member of the genus Lilium, belonging to the Liliaceae family is one of the most important commercial pot and cut flower species and one of the three major bulb crops in the commercial market because of its large, colorful and fascinating flowers. Lily hybrids are the most economically important plants with varied flowers. Hybrid Eastern lily (Lilium oriental hybrid ‘Casablanca’) is a perennial bulbous plant that its propagation by bulb in natural condition is time-consuming, so produces 1–2 bulblets per bulb scale in one years’ time which is not sufficient for large scale cultivation of this plant. One of the most important and best methods for vegetative propagation and breeding of lilies is in vitro bulb scale culture. In vitro adventitious bud regeneration from scales of Lilium rely on many factors like cytokinin and auxin concentrations such as BA and NAA. The successful use of tissue culture techniques for rapid propagation of some species of the genus Lilium including L. ledebourii, L. orientalis, L. longiflorum, L. japonicum, L. speciosum, L. concolor, L. nepalense, L. regale, L. oriental hybrid, L. Asiatic hybrid has been reported. The purpose of the current study was to evaluate the effect of different concentrations of BA and NAA on in vitro proliferation of Lilium oriental hybrid ‘Casablanca’ using bulb scale as explant to establish a suitable protocol.
 Effect of various concentrations of 6-benzyle adenine (BA; 0, 0.5, 1 and 2 mg l–1) and ɑ-naphtaleneacetic acid (NAA; 0, 0.1, 0.2 and 0.4 mg l–1) were evaluated on in vitro proliferation of L. ‘Oriental’. Bulb scale as explant and MS basal medium as culture medium were used. Activated charcoal was applied to inhibit the browning of the culture medium and explant. The experiments were conducted in completely randomized design (CRD). The 16 treatments were applied, each treatment had 4 replications and each replication had 4 individuals. Therefore, in these experiments, a total of 192 bulbs were used. Traits including total plantlets fresh weight, leaf length, leaf number, bulblet weight, bulblet diameter, bulblet number, survival percentage, root length and root number related to in vitro proliferation were measured. All the statistical analyses were done by using SAS and Tukey’s test. Arcsin software was used for changing percent data.
 The interaction effect of BA and NAA was significant for all measured traits. Results showed that the maximum number of bulblet (8.66) and root (5.36) were obtained in culture medium enriched with 0.5 mg l–1 BA together with 0.4 mg l–1 NAA. Culture media supplemented with 0.5 mg l–1 BA together with 0.2 mg l–1 NAA and 1 mg l–1 BA together with 0.1 mg l–1 NAA with induction of 7.33 bulblets per explant were suitable media. The largest number of leaf (4.33) was measured in culture medium containing 1 mg l–1 BA together with 0.1 mg l–1 NAA. The highest bulblet weight was measured in culture medium supplemented with 1 mg l–1 BA along with 0.2 mg l–1 NAA. The greatest survival rate (100%) was observed in medium enriched with 0.5 mg l–1 BA together with 0.1 mg l–1 NAA. Survival rate (90%) in explants treated with 2 mg l–1 BA along with 0.4 mg l–1 NAA was high. Obtained results revealed that the presence of both BA and NAA in culture media for enhancement of most traits is necessary and critical. Plantlets were transferred to a growing medium containing cocopeat, peat moss and perlite in identical proportion for acclimatization following proliferation. Approximately, 90% of regenerated plantlets survived and were morphologically similar to the mother stocks. This study will help the producers and breeders for commercial and improvement purposes. The effective role of the simultaneous presence of both auxin and cytokinin in the culture medium in effective organogenesis was shown in the present study. Similar findings were reported for some lilies such as L. ledebourii (Baker) Bioss., L. longiflorum and L. regale. Auxin was effective in stimulating bulb production and growth of the aerial part of the eastern lily, and its presence along with cytokinin is essential for leaf induction. Some studies have reported similar results. The type and optimal concentration of plant growth regulators (PGRs) in the culture medium for suitable in vitro propagation varies in different species. Genetic variations (species type), differences in the amount of endogenous production of PGRs and their interaction with each other are among some reasons for this difference. The proper ratio of auxin and cytokinin in the culture medium is effective for inducing cell division, cell differentiation, organogenesis and finally for achieving a complete plant. Root production with appropriate quantity and quality leads to the suitable survival of seedlings resulting from the growth of cultured explants under in vitro conditions and adapted plants. Current study showed that the presence of both BA and NAA is better than the presence of one of these two PGRs for induction and growth of root. Some similar findings were reported, however in most studies, the presence of auxin as individual PGR has been found to be more suitable for root induction.

Orchis muscula L. is one of the worthwhile ornamental, medicinal, native and endangered species in Iran. In this research, plant tissue culture method has been used to propagate this valuable plant. For in vitro organogenesis, protocorm explants were surface sterilized and then cultured on different shoot and root induction media based on OrchiMax (OM) medium, supplemented with plant growth regulators with different concentrations alone or in combination. Based on results, the most shoot induction (100%) was obtained from protocorm explants cultured on medium containing 2 mg L-1 BAP and 0.5 mg L-1 NAA and the highest root induction (80%) was belonged to the treatment of IAA at 1 mgL-1.

The extinction threatens some native orchids which has economic value. Artificial seed technology can prevent the extinction of these plants and can be suitable for producing and exporting of product. In this study, protocorms of Dactylorhiza umbrosa were encapsulated. The effects of three temperature treatments 4, 20, and 25°C were investigated during three periods 30, 60, and 90-days of artificial seed storage. Percentage of artificial seed germination during storage conditions, after storage, total percentage of germination and number of days required for germination incidence, were investigated. The highest germination percent during storage conditions was related to 90-days storage at 4 and 25 ° C, and the lowest obtained after 30 days at 4 ° C, which did not show any germination. The highest germination percentage of artificial seeds after storage and cultivation was observed in 60 days stored seeds at 4 °C. The highest total percentage of germination was obtained after 60 and 90 days storage at 20 and 4 °C, respectively. The highest number of days befor germination incidence was observed after 60 days of storage. The best treatment for storage of artificial seeds was 4 °C for 30 and 60 days, which showed the lowest percentage of germination during storage conditions and the highest percentage of germination after culturing.

The walnut family (Juglandaceae) consists of approximately 60 species of deciduous trees is native of the American continents, Europe, and Asia. Pecan (Carya illinoensis) is belonged to the Juglandaceae family and is one of the most valuable nut products all over the world. Embryo culture techniques for plant breeding as well as basic studies in physiology and biochemistry are widely used. The low percentage of germination and the long propagation cycle and the need for stratification treatments from three to six months are the most important barriers to the development of high yielding cultivars through hybridization. Plant regeneration methods from embryo culture in vitro allows overcoming the barriers of hybridization, as well as obtaining higher and faster multiplication rate of plants of an elite genotype.

In this experiment, an adapted native genotype of pecan in Gorgan city, Golestan province, Iran was selected. The mature fruits were harvested after five months of pollination. They were immediately transferred to the laboratory. For cold pretreatment, nuts packed in a paper bag and stored in 4-5ºC for 15 days. The effect of two types of culture medium, growth regulators and seed pretreatment (15 days at 4-5 °C) on germination of mature embryos of pecan has been determined. Murashige and Skoog (MS) and Woody Plant Medium (WPM) and IBA (0 and 1 mgl-1), BAP (0, 1 and 2 mgl-1) and GA3 (0 and 1 mgl-1) media were used to embryo rescue evaluation. The data obtained were statistically analyzed in completely randomized block design (RCBD). Each treatment was replicated at least third, and each replicate consisted of two zygotic embryos. Means of germination period, percent of seed germination, root and shoot length and leaf number in different media and various PGRs combination were compared based on LSD at p ≤0.05.

The results showed, although cold pretreatment for 15 days had no effect on germination period, root length and number of leave but also, effect on germination percentage and shoot length. There are some different hypothesis about the effect of cold pretreatment on embryo germination between researchers. Some researchers believed that, there is low efficiency in embryo germination in lack of cold pretreatment and GA3. Cold pretreatment or GA3 reduce the ABA level and promote embryos germination. The others reported poor germination for somatic embryos when they treated with GA3 and cold pretreatments. Pearce et al. (1987) reported that GA3 and substrate of GA3 can be increased during the chilling process as ABA levels decrease. Furthermore, application of exogenous GA3 induces germination. Tang et al. (2000) reported that somatic embryos germination poorly happened in cold condition and addition of GA3 did not change the poor germination. Kaur et al. (2006) and Peyghamzadeh and Kazemitabar (2010), reported that the embryo germination in Juglans regia L. was higher when GA3 and cold pretreatments were simultaneously applied as compared to those when applied separately. In this experiment, media has no effect on embryo germination period but, could effect on other parameters. As the results showed, MS media showed the maximum percentage of germination, root and shoot length and number of leave in both condition (with and without cold pretreatment). In this experiment root length of germinated pecan embryo was higher in MS medium. Mapelli et al. (2001) reported that seed germination resulted in marked changes in the metabolism of free amino acids in walnut cotyledons. About 52% of the total free amino acids in one-month-old seedlings was present in the cotyledons and about 26% was in the taproot. The concentration of free amino acids in the taproot was similar to that in the embryonic axis, and greater than that in the cotyledons. T11 (1mg/l-1 IBA, 1mg/l-1BAP and 1mg/l-1 GA3) and T12 (1mg/l-1 IBA, 2mg/l-1BAP and 1mg/l-1 GA3) treatments were the highest in germination percentages in both treatment (with and without cold pretreatment). There was no significant differences between 1 mgl-1 and 2 mgl-1 of BAP.

Pecan as like walnut, is considered to be one of the most recalcitrant species in vitro. It is necessary to determine the optimal culture conditions to establish it for shortening time in seed propagation. This seedling could be applied as primary material for breeding programs, grafting and physiology study. The best growth of micro plant achieved in MS medium with 1 mgl-1IBA, 1 mgl-1GA3 and 2 mgl-1BAP.

Epipactis veratrifolia is a terrestrialorchid that belongs to thetemperate zones of Iran with great breeding potentials for ornamental usages. Although in vitro propagation of this orchid has been previously performedbut investigation on factors influencing its in-vitro growth and photosynthesis is still needed. In the current study, three-leaflet seedlings were cultured in closed vessels containing modified Fast liquid medium together with sterilized perlite (for sample establishment). To remove CO2, vials containing saturated KOH solution as CO2 absorbent and to increase CO2 concentrations, vials containing 3 mL CO2-releasing solutions (3 M sodium bicarbonate and sodium carbonate) were placed into the culture vessels. The experiment was carried out based on a completely randomized design with three replications and three samples per each replicate. Growth parameters including fresh and dry weights of shoot, stem height and root length together with transpiration rate and maximum quantum yield of photosystem II efficiency (QYmax) of the samples were measured. The highest values for shoot height, shoot dry and fresh weights and root length were observed in CO2-increased treatments. Removing CO2 from the atmosphere of culture vessels caused a dramatic decrease in QYmax of plantlets in vitro. Meanwhile, following desiccation, higher transpiration rate was detected in control and CO2-increased treatments, while removing CO2 decelerate transpiration rate of the plantlets. In conclusion, with increasing CO2 in vitro we can promote photosynthetic efficiency of Epipactis veratrifolia and as a result enhance growth of plantlets in vitro.

This research was conducted in two stages to optimize in vitro propagation of Persian wild paeony. Stage 1: effect of BAP (0, 0.5, 1 and 2 mg.l-1), NAA (0, 0.1, 0.2 and 0.3 mg.l-1) and GA3 (0 and 0.5 mg.l-1) on apical bud ofPaeonia mascula and stage 2: effect of BAP (0, 0.5, 1 and 2 mg.l-1) and NAA (0, 0.25, 0.5 and 1 mg.l-1) were studied on embryonic callus induction on young leaves of Paonia mascula. At stage 1: the highest shoots number (5.38) was obtained on media containing 1 mg.l-1 BAP + 0.1 mg.l-1 NAA+ 0.5 mg.l-1 GA3. The results showed that increasing BAP level caused decreasing of shoot length. At stage 2: maximum of compact embryonic callus (17.93%) and somatic embryo number per explant (7.11) were obtained on 2 mg.l-1 BAP and 0.5mg.l-1 NAA. For callus proliferation, calli were maintained on MS medium containing 2 mg.l-1 BAP, 0.1 mg.l-1 NAA and 0.5 mg.l-1 GA3. Maximum rooting percentage (70.51%) and root number (0.0.02) were obtained with 0.25 mg.l-1 NAA.

To evaluate the effects of sucrose and and ultrasound on Chelcheragh lily (Lilium ledebourii Boiss.) under in-vitro condition, an experiment was conducted in a factorial experiment based on completely randomized design with four replications in tissue culture laboratory of Department of Horticultural Science in Mohageg Ardebili University in 2014. Treatments in this study were sucrose in 3 levels (30, 45 and 60 g L-1) and ultrasound with frequency of 35 kHz in 5 levels (0, 5, 10, 20 and 30 seconds) on the 2 types of explants (bulblet and scale). In bulb explants, the results showed that the highest fresh weight, bulbs and roots weight, root and plantlet length was obtained from 60 g L-1 sucrose + 10 seconds ultrasound, but the least amount of total phenolic content was observed from this treatment combination. The highest total phenolic content was obtained from 60 g L-1 sucrose + 30 seconds ultrasound. All levels of ultrasound had positive effect on number of scales, number of leafs and leaf length, but reduced the amount of chlorophyll. The highest number of roots and number of rooted bulbs was observed in MS medium containing 30 g L-1 sucrose. In both explants, MS medium containing 45 g L-1 sucrose was effected in increase of length of root. In second experiment, ultrasound destroyed the scale explants.

Fritillaria imperialis L. is an ornamental and medicinal plant native to mountainous regions of Iran. This plant genetic resource is in danger of extinction, Because of grazing livestock and pest outbreaks. Therefore, micro propagation of Fritillaria through in vitro regeneration is essential for conservation and commercial production. Thymol and Carvacrol are one of the main essential oil compounds in family Lamiaceae.
Material and Fritillariaimperialis L. bulbs in dormancy stage obtained from mountainous regions of Lorestan in Iran and were placed in cold room at +4 °C for 4-6 weeks. Then, Bulbs were surface-sterilized with 70% ethanol for 45 s followed by immersion in 5% (v/v) NaOCl solution for 20 min with gentle agitation, and then rinsed three times in sterile double distilled water. Present study was conducted in two separate experiments. In first experiment, effect of different concentration of Thymol and Carvacrol and in second experiment, different concentration of NAA and BA on in vitro characteristics of Fritillaria was evaluated. Explants (1× 1 cm) prepared from the lower third of scales with basal plate and were placed in MS basal medium supplemented with different concentrations of Thymol (50, 100, 150 and 300 ppm), Carvacrol (10, 100, 500 and 100 ppm), BA (1, 2 and 4 mg/l) and NAA (1, 2 and 4 mg/l).All cultures were incubated in a growth chamber at 24±2°C, and a photosynthetic photon flux of 40-60 μmol m–2 s–1 was provided by cool white fluorescent lamps with a 16-h photoperiod. This experiment wascarried out in completely randomized designs with fivereplications. Analysis of variance showed that Thymol and Carvacrol were not effective on number of new bulblets but had significant effects on bulb diameter, number and length or roots, number and length leaves and callus induction and diameter of callus obtained from scales (P< 0.05). The highest rate (3 bulblets) of bulblets formation was obtained fromMS medium supplemented with 50 ppm Thymol that showed significantly difference from other treatments. Medium containing 10 ppm Carvacrol gave the highest Bulblet formation (2.5 bulblets) between Carvacrol treatments. Investigation of rooting was done by assessment of the number and length of roots. Mean comparison of the effect of cultivar type on root number showed that the largest number of roots per explant was obtained fromMS medium containing 50 ppm Thymol. Lowest number of roots observed in mediums supplemented with 300 ppm Thymol and 100 ppm Carvacrol. The best medium for increasing the root length per explant (10.90 cm) was MS medium supplemented with 100 ppm Carvacrol, while the least increasing in root length per explant observed from culture mediums contained 300 ppm Thymol and 100 ppm Carvacrol. Also, the largest number of leave formation obtained from culture medium supplemented with 50 ppm Thymol that significantly higher than other treatments. Statistical analysis (ANOVA) of the data showed that high frequency callus induction and formation occurred in MS mediums contained 50, 100 and 150 ppm Thymol and 10 ppm Carvacrol and culture mediums supplemented with 300 ppm Thymol and 1000 ppm Carvacrol showed least callus induction. In contrast, largest callus diameter observed in culture mediums supplemented with 300 ppm Thymol and 500, 100 ppm Carvacrol.
Statistical analysis of results showed that different concentrations of BA and NAA had significant effects on bulblets number and bulblets diameter (P

A standard in vitro mass propagation protocol for development and propagation of valuable flowers such as Anthurium is one of the major procedures and goals in flower industry. Towards developing a commercialized propagation recipe for A. andraeanum cv. Pink, effects of BA and NAA were investigated on regeneration, proliferation and in vitro mass propagation efficiency of cultivar Pink, as well as quantitative and qualitative characteristics. The lateral buds were established in 1/2MS contains different concentrations of BA (0, 0.1, 0.3 mg/l) and NAA (0, 0.04, 0.07, 0.23 mg/l). The longest shoots (average of 23.46 mm), was obtained in 1/2MS containing 0.23 mg/l NAA .The highest number of buds were produced in medium containing 0.23 NAA plus 0.3mg/l BA. The highest root number was emerged in 0.23 mg/l NAA. The highest percentage of regeneration rate obtained in the media containing 0.23mg/l NAA 0.3mg/l BA compared to the control. Obtained protocol can be simply used for in vitro mass propagation of healthy Anthurium plants.

The loganberry (Rubus × loganobaccus), belonging to the Rosaceae family and Rubus genus, gives fruits like strawberry. The fruit is red and very sour and this rare species is one of the cultivars of red raspberry. Loganberry is not native to Iran and currently is propagated through in vitro plant tissue culture. So far, phytochemical research has not been conducted on loganberry. In present study, the leaf extract of loganberry, propagated through in vitro culture, was investigated phytochemically. The extract of loganberry was purified and the obtained natural product was elucidated by NMR spectroscopy techniques. In this study, a flavonoid, namely hesperetin, was isolated from the leaves of the loganberry. This structure was elucidated by spectral methods (1H-NMR, 13C-NMR, IR and dept 135).

Lily (Lilium spp.) is a genus of herbaceous flowering plants, which consisted of many beautiful ornamental species with large prominent flowers. Most species are native to the Northern hemisphere temperate, though their range extends into the Northern subtropics. Some specific hybrids of Lilium spp. have been developed as cut flower in controlled conditions and in some cases can be grown as pot plant. Propagation rate of lily in natural clonal propagation methods is very low and one year produces of 1-2 bulblets per bulb scale. There is also possibility of disease transmission; so that, tissue culture techniques has provided an efficient method for its micropropagation.
In this study, two separate experiments under In vitro conditions the bulblet regeneration from thin cell layer (TCL) explants of Lilium spp. was investigated. In the first experiment, after two months the effect of TCL explants with 1, 3 and 5 mm thickness on MS medium containing 1 mg/l BA, 2ip and kin in combination with 0.5 mg/l NAA on regeneration parameters were assayed. In the second experiment to determine the effect of cultivar and cytokinin types, 3mm thickness TCL explants of five cultivars (Robina, Donato, Nymph, Lessoto and Roxana) were tested on MS medium containing different plant growth regulator (PGR) compounds including BA, kin, 2ip and TDZ at concentration of 1 mg/l in combination with 0.5 mg/l NAA. The regeneration parameters were assayed after four months. In all experiments, the medium was adjusted to pH 5.8 and autoclaved at 121°C for 15 min. All cultures were incubated at 25 ± 2°C with a 16 h photoperiod under cool white flourescent lights (30 µmol/m2).
According to the first experiment results, plant growth regulator of BA in all of surveyed parameters except root number was better than other PGRs and explants with 3 mm thickness was the best in all of parameters. The interaction of PGR and explants was significant, however maximum bulblet regeneration was observed in TCL explants with 3 mm thickness in all of PGR treatments (100%). While 1 mm thickness TCL in 2ip and 1 and 5 mm thickness TCL in Kin had the least regeneration percentage. Results revealed that the interaction of explants and medium is a key factor for suitable establishment, regeneration and growth of TCLs. Bulb dormancy is one of the limiting factors in regeneration of bulbous crop species. It seems under In vitro condition explants size and PGR combination of media especially cytokinin affected on breaking of dormancy. Maximum number of leaves and dry weight of bulblets in medium containing BA was significantly higher compared with other treatments. Most of studies confirmed the positive effect of BA on regeneration of lily. The function of cytokinin in plant promoted cell division and differentiation, which lead to growth and maintaining cells in meristematic status.
Result of second experiment showed that cultivar was one of the effective factors on regeneration trait. Oriental lily cultivar "Roxana" had the highest number of roots, bulblets, dry weight and length of plantlets and "Nymph" cultivar showed the lowest percentage of regeneration, dry weight, length of plantlets and rooting obtained. In all of cultivars BA induced more organogenesis percentage and plantlet dry weight, while TDZ induced more rooting percentage.The interaction of cultivar and PGR treatments on percentage of regenerated bulblets and rooting were significant. "Nymph" cultivar had minimum percentage of regeneration and rooting in medium containing TDZ and Kin. Furthermore, "Roxana" cultivar in medium containing BA showed the best dry weight comparison to other treatments.
Conclusion Lily has widely used in the floral industry as a cut flower or potted plant. In recent years, tissue culture was developed as reliable and highly effective method to overcome its limitations of vegetative propagation. The most advantage of this method is high multiplication rate and disease free propagation. In this study, bulblet regeneration of lilium Spp. from TCL explants under in vitro condition was considered as a highly efficient procedure for its micropropagation. With optimization of TCL system some parameters such as exogenously applied plant growth regulators, cultivar, explants types were investigated. Favorable conditions for bulblet regeneration were achieved with 3 mm thickness TCLs in MS medium containing 1 mg/l BA with 0.5 mg/l NAA. This protocol can be used for rapid micropropagation of many cultivars.

For improvement asymbiotic seed germination media of temperate terrestrial orchid Epipactis veratrifolia , 12 different concentrations of the carbohydrates (fructose, glucose, sucrose and two combination of fructose with sucrose ) were assessed on the seed germination and protocorm development, in the presence (2g L−1) and absence of peptone in Fast medium. Results revealed significant differences between treatments on seed germination percentage and protocorm growth. Carbohydrate treatments; H12 (3.5g/l fructose g/l sucrose), H8(10 g/l sucrose), H2 (20 g/l glucose), and H11 (5 g/l fructose g/l sucrose) had significant effect on seed germination percentage. H11and H12 was the best medium for protocorm growth. H8P2(10 g/l sucrose g/l peptone), H2P2(20 g/l glucose g/l peptone) and H12P2(3.5g/l fructose g/l sucrose g/l peptone) were the best for seed germination respectively with 79/6%, 74/6% and 71/9% seed germination percentage. H10P2 (30 sucrose g/l g/l peptone) with 17/3mm growth, significantly was the best for protocorm growth. Therefore kind and concentration of carbohydrate and presence of organic nitrogen (peptone) influence asymbiotic seed germination percentage and protocorm growth and could improved both of them. This finding revealing that it is possible to improvement asymbiotic seed germination of this species orchid with combination of mono and disaccharides and organic nitrogen.

Echinacea is one of the most important medicinal plants and an important source of cichoric acid. In this research, callus induction, indirect shoot regeneration, and shoot rooting of Echinacea angustifolia DC. were investigated. The effects of explant (leaf, petiole and root) and NAA concentrations (0.5, 1 and 1.5 mg l-1) on callus induction as well as the effects of explant (leaf, petiole and root) and BAP concentrations (3, 4 and 5 mg l-1) on the indirect shoot regeneration of callus were evaluated in two separated experiments. The study was conducted as factorial in a completely randomized design (CRD). The effects of IBA concentrations (0.5, 1, 1.5 and 2 mg l-1) on shoot rooting were also investigated in a completely randomized design. The effects of the factors studied on the callogenesis were not significant while these factors had significant effects on the shoot regeneration and rooting at 1% probability level. In general, the results indicated that the highest indirect shoot regeneration (72%) was obtained from the leaf explant and by the use of 4 mgl-1 BAP. In addition, the highest shoot rooting (38% and 40%) was obtained using 1 and 1.5 mgl-1 IBA. The good potential of this species for indirect shoot regeneration was shown in this research. These results can be used in micropropagation programs, gene transfer, and transgenic plants production.

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.

In this research, complete micropropagation protocol of GF677 has been described by indirect organogenesis from embryo culture. In order to break the chilling requirement, mature seeds after disinfection were treated with 250 mg-l of sterile solution of GA3 for different periods of time (0, 12, 24, 48, 96 h). Then, embryo explants separated carefully from the seeds and for callus induction were cultured in factorial experiment on Murashige and Skoog medium (MS) supplemented with different growth regulator treatments. The results showed, only in the presence of 2,4-D and BA, GA3 treatment increased callus induction of explants. Calluses were transferred on medium containing BA (0.0, 1.0, 2.0 and 4.0 mg-l) with 0.25 mg-l NAA and 0.25 mg-l IBA. The highest percentage rate of regeneration (90%) and the highest shoot number, (16.60) per explant were obtained in the presence of 2.0 mg-l BA. Seeds treatment with GA3 at this stage reduced regeneration of shoots from callus. For rooting of regenerated shoots, the effect of three types of auxin IBA, NAA and IAA in different concentrations (0.0, 0.5, 1.0, 2.0, 4.0 and 8.0 mg-l) on 1/2MS medium were studied. The highest rooting index was observed in the presence of IBA. Rooted plantlets after transferred to the soil mixture (peat, perlite and sand, 2: 2: 1) acclimatized with 85% success in greenhouse condition.

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.

In order to investigate the effect of two spermidine concentrations (0.5 and 1 µ M) on in vitro gynogenesis of six onion accessions from Khorasan province this research was carried out. Results showed that the highest and lowest percentage of gynogenesis rate was belonged to M2 and M5 media, respectively. The highest (1.41) and lowest (0.33) percentage of gynogenic embryo were belonged to Roshnavand-e- Birjand and Sefid-e- Naishabur accessions, respectively. The highest percentage of plant regeneration was belonged to Roshnavand-e- Birjand accession in M2 medium culture. Ashkhane-e- Bojnourd and Dargaz accessions had the highest and lowest percentage of plant survival rate (81.25 and 49.8, respectively). Therefore, spermidine in two concentrations of 0.5 and 1 µ M, without other plant growth regulators, had no positive effect on onion gynogenesis and should not be used instead of 2,4-D and BA. Combination of spermidine with other plant hormones could induce gynogenesis in onion.

To protect and multiply important and rare plant resources، in vitro culture serves as a more efficient alternative to traditional propagation approaches. Levisticum officinale Koch. a member of Apiaceae is an important، endangered and neglected species in Iran، which has been shown to have diuretic، spasmolytic and carminative effects. In order to supply enough plant materials for micro-propagation of this herb and study effects of different methods of disinfection and stratification on in vitro seed germination، a factorial experiment laid out in a completely randomized design was set out to establish sterile plants out of seed culture. It was concluded that a pre-chilling treatment for 3 months resulted in maximum percent of germination (92%) and the largest germination rate. The best superficial sterilization protocol was proofed to be soaking in 70% (v: v) ethanol for 30 s and then، using of 2% (v: v) dilution of NaOCl for 15 min، followed by 3 rinses in sterile distilled water.