پژمان آزادی

Susan-e-Chelcharagh (Lilium ledebourii) is a rare native plant in Iran, primarily found in the elevated terrains of the Alborz region. This study explores the potential for crossbreeding L. ledebourii with commercially available lily varieties. Initially, the study compares the cytogenetic profile of L. ledebourii with several commercial cultivars. Subsequently, the feasibility of hybridization between L. ledebourii and a subset of commercial lily cultivars were analyzed. The study also examines the viability of in ovulo culture in maternal genotypes of Brindisi and Paradero cultivars when cross-pollinated with L. ledebourii pollen. Moreover, the effects of different time intervals (10, 20, and 30 days after-pollination) and NAA concentrations (ranging from 0 mg/L to 2.0 mg/L) on ovule development within MS medium were considered. The findings reveal that L. ledebourii is a diploid plant with functional pistils and pollen grains. However, when used as the maternal parent in crossings with commercial cultivars, all pistils undergo abscission, preventing fruit set. Additionally, fruit set is conspicuously absent when L. ledebourii serves as the paternal parent in crosses with the Companion, Fangio, and Arvandrud cultivars, and fruit formation is transient in other examined cultivars, disappearing completely by the eighth week post-pollination. The results of the in ovulo culture assays highlight the significant influence of isolation time and NAA concentration on ovule viability and enlargement. Notably, in the Brindisi cultivar, ovule germination becomes apparent when isolated 30 days after-pollination and supplemented with 0.1 mg/L NAA. In conclusion, interspecies hybridization involving L. ledebourii as the pollinating parent and select commercial cultivars remains possible by skillfully managing pre- and post-fertilization barriers through using optimal cultural media.

Camelina (Camelina sativa), an oilseed plant in the Brassicaceae family, is recognized as a significant crop both biologically and industrially, with recent attention shifting towards its potential as a biofuel source. The close genetic resemblance between Arabidopsis thaliana and C. sativa has sparked interest among researchers in manipulating oleic acid levels through microRNAs and the gene sequences FAE1-A and FAD2-A. A recent bioinformatics study focused on these genes in camelina revealed that the FAE1-A protein is hydrophobic, while FAD2-A is hydrophilic. Structural analysis indicated GMQE values of 0.88% for FAE1-A and 0.93% for FAD2-A. The FAE1-A protein’s secondary structure comprises 49% helix, 11% beta-sheet, 41% coil, and 9% membrane content with a confidence level of 79.2%. Similarly, the secondary structure of the FAD2-A consists of 43% helix, 12% beta-sheet, 45% coil, and 30% membrane content with a confidence level of 79.8%. Codon preference patterns were explored using the Sequence Manipulation Suite database to understand the relationship between codons and gene performance. Furthermore, analysis of FAE1-A and FAD2-A gene expression showed peak expression levels in developing seeds approximately 20 days after pollination. Further investigation into these structures promises to enhance our understanding of fat biosynthesis, thereby improving oil quality in C. sativa.

Chrysanthemums are known as one of the top three flowers in the floriculture industry. This ornamental plant is traditionally propagated by cuttings. The use of in vitro subculture is expanding for plants like chrysanthemums that are propagated by asexual methods. This research evaluated the effect of plant growth regulators on callus induction and regeneration in three chrysanthemum cultivars including ‘Bonfire Yellow’, ‘Rambla’, and ‘Bella Rosa’. For callus generation, the leaf explants were cultivated in the Murashige and Skoog (MS) culture medium modified with with benzyl adenine (1, 2, or 3 mg/L BA) and naphthaleneacetic acid (0.5 or 1 mg/L NAA) in a factorial experiment based on a completely randomized design. The highest rate of regeneration of ‘Bonfire Yellow’(31.25%), ‘Rambla’(25%), and ‘Bella Rosa’ (31.25%) were obtained from the culture media containing 2 mg/L BA + 1 mg/L NAA, 1 mg/L BA + 1 mg/L NAA, and 3 mg/L BA + 0.5 mg/L NAA, respectively.

Improving the quality and increasing flowering is one of the goals of growing bulbous flowers. It seems that plant growth stimulants depending on the type and concentration used can be effective in achieving this goal. This experiment was performed to investigate the effects of type and concentration of three growth stimuli on flowering and corm production of freesia based on a completely randomized design with three replications. Experimental treatments included three types of growth stimulants (bioradicant, Ecormon and Futop) at three levels of 0.5/1000, 1/1000 and 1.5/1000. No foliar application was considered as control treatment. The results showed that with foliar application of 1/1000 Futop, leaf length increased by 8.9% and 14.5% and leaf width by 9.8% and 5.2%, respectively, compared to the control (no foliar application). Also, in plants treated with bioradicant growth stimulant at the concentration of 1.5/1000, leaf area was higher than other treatments. The use of growth stimulants reduced flowering time. In all three growth stimuli used, an increase in flowering stem length was observed compared to the control. The use of growth stimulants in all 9 treatments was effective in increasing the number of peduncles. Using the highest concentration of growth stimulants used in this experiment, ie concentration of 1.5/1000, the diameter of the peduncle in bioradicant, ecormon and futop treatments reached 21.7, 20.6 and 23.5 mm, respectively. Bioradicant treatment at the concentration of 1.5 /1000 resulted in 2.3-fold increase in the weight of the peduncle. Total chlorophyll increased from 23.22 μg/fw in no foliar application to 31.89, 29.80 and 31.47 μg/fw at the concentration of 1.5/1000 growth stimulants of bioradicant, ecormon and futop. According to the results, different concentrations of growth stimulants caused a change in the percentage of nitrogen and phosphorus in freesia leaves. Accumulation of phosphorus in leaves treated with ecormon was more than the other growth stimulants used. In general, among the three growth stimulants used, bioradicant at the concentrations of 1 and 1.5/1000 had greater effect on the vegetative and reproductive quality of freesia.

The formation of a flowering stem in Lilium requires the production of commercial bulblet and its breaking dormancy. Three experiments, each as a completely randomized design based on a factorial experiment, were conducted. The first was to evaluate the effects of cultivar and naphthalene acetic acid (NAA) (100 mg/L) on bulblet production rate in scales. In the second, with the aim of investigating the effect of medium culture and bulblet size on bulblet size-changing percentage, bulblets were classified into three sizes according to circumference and weight, then cultivated in two peat:perlite (70:30) and sandy medium. Six commercial hybrids were used in these two experiments. In the third, the effect of bulblet size, chilling, heating, and gibberellic acid on dormancy breaking of the top three cultivars were investigated. In the first experiment, NAA did not have a positive effect on bulblet production, and the highest production rate (4.1 bulblets per scale) was seen in not treated scales with NAA in ‘pinnacle’. According to the results of the second, the largest bulblet and peat:perlite medium showed the best results. The highest bulblet circumference-changing percentage (99.7%) and the highest weight-changing percentage (180.56%) were obtained in the largest size of ‘Pinnacle’ bulblet in peat:perlite medium. According to the results of the third, more than 60% of gibberellic acid-treated bulblets in ‘Pinnacle’ and ‘Eyeliner’ flowered, and the largest bud (8.57 cm) produced by the largest bulblet in ‘Pinnacle’. Therefore, ‘Pinnacle’ and ‘Eyeliner’ were introduced as suitable cultivars for production and gibberellic acid as a chilling replacement for dormancy breaking.

The present study has been designed and executed to determine the best growth-regulating compound for callus induction as well as to specify the optimum dose of gamma irradiation in carnation cultivars (Tabasco, Nobless, Cameron, Tabor, Eskimo, and Mariposa). In this experiment, an MS culture medium was used to evaluate the various levels of growth regulator concentrations including NAA in four levels (0, 0.5, 1, and 2 mgl-1), 2,4-D in five levels (0, 0.5, 1, 2, and 3 mgl-1), and BA in two levels (0.5 and 1 mgl-1). Irradiating the callus of leaf explants was carried out three weeks after cultivation at 0, 15, 25, 35, 45, and 55-gray doses-to determine the optimum dose of gamma radiation. The analysis of data and illustration of graphs were carried out via Excel software and according to the obtained results, the radiation level that killed 50% of the calluses was selected as the optimum dose for further experiments. The results have indicated that all main effects and the interaction effects regarding the characteristics of callogenesis percentage and callus volume were significant at a probability level of 1%. Means were grouped using Duncan's multiple range test, revealing that the highest level of callus induction was in Eskimo cultivar with a 73% overall mean. Overall, the results indicate that 2 mgl-1 2,4-D, 0.5 mgl-1 BA is the best regulatory compound for callogenesis in carnation cultivars. Moreover, it was found that on average, the 25-gray dose leads to suitable results in the callus explants of all cultivars.

In the study, to survey the growth, yield, and physiological responses of valerian (Valeriana officinalis L.) using different concentrations of urea fertilizer (0, 30, 60, 90, and 150 kg.ha-1) and nitroxin and phosphate Barvar-2 bio-fertilizers, an experiment was conducted in medicinal Plants farm, Tehran, as a factorial in a randomized complete block design with three replications in the 2019-2020 crop year. Seedling cultivation was carried out in mid-May and urea fertilizer treatment was performed in two stages. To measure root and biological yield, one square meter was harvested from each plot and the dry weight of shoot and root was measured and the sum of these two was calculated as biological yield. Root and rhizome essential oils were extracted by water distillation (Clevenger) and valerenic acid by high performance liquid chromatography (HPLC). Essential oil yield was also calculated based on the essential oil percentage and dry root yield. The average root diameter varied from 7.97 (90 kg urea per ha) to 5.14 mm (control). The highest root yield was obtained in 150 and 90 kg urea.ha-1 along with inoculation with nitroxin and phosphate Barvar-2 (4.88 and 5.1 ton.ha-1, respectively). Application of 90 kg urea.ha-1 along with nitroxin resulted in increased chlorophyll b and total content. Increasing the urea concentration decreased essential oil content and increased essential oil yield. Essential oil content ranged from 0.93 (90 kg urea.ha-1 with nitroxin and phosphate Barvar-2) to 1.94% (no-application of fertilizers). The highest valerenic acid content (0.46% of the extract) and valerenic acid yield (220.27 mg.ha-1) were observed using 30 kg of urea with phosphate Barvar-2 and no application of nitroxin. In general, application of nitroxin and phosphate Barvar-2 bio-fertilizers along with 90 kg urea.ha-1 by improving growth, yield, and physiological traits led to the production of acceptable essential oil and valerenic acid yield.

Caladium is an attractive ornamental plant that is very popular on the ornamental plant market due to its shape, color and leaf variety. This plant is propagated by dividing the underground stem traditionally, which does not meet the needs of the market due to low speed propagation. This study was designed and conducted to propagate this ornamental plant through tissue culture. After disinfection, leaf explants were placed on MS medium with different concentration of BAP (0.5, 1, 2 and 3 mg.l-1) and NAA (0.2 and 0.5 mg.l-1). After 4 weeks, the regenerated plants were transferred to proliferation medium including MS consisting of BAP (1, 2, 3 and 4 mg.l-1) and NAA (0.01, 0.1 and 0.5 mg.l-1). After the proliferation stage, the plants were transferred to the rooting medium containing IBA (0.1, 0.5 and 1 mg.l-1). Based on the results, the highest percentage of direct regeneration was obtained from explant leaf in medium containing 2 mg.l-1 BAP and 0.2 mg.l-1 NAA. The highest number of shoot was observed in MS medium with 3 mg.l-1 BAP and 0.1 mg.l-1 NAA and the highest shoot height in 4 mg.l-1 BAP with 0.1 and 0.5 mg.l-1 NAA. The plantlets were rooted in 0.5 mg.l-1 IBA as optimal concentration. Finally, the rooted plants have been successfully adapted to the greenhouse (With an average of 25±2°C and a relative humidity of 70 to 80%).

The genetic improvement of carnation (Dianthus Caryophyllus), as one of the most important cut flowers in the world, is of utmost significance. For this purpose, the use of in vitro breeding techniques is very important. One of the most crucial in vitro breeding needs of carnation is accessibility to a regeneration method with maximum productivity and minimum genetic modification probability. The present study aimed to determine an optimal and fast method for the direct regeneration of carnation cultivars by employing various regulators and adenine hemi-sulfate for the first time in these cultivars.

For the determination of the optimal level of the culture medium compositions toward the direct regeneration of carnation cultivars (Skimo, Tibor, and Labret), an experiment with a completely randomized design was designed and implemented in an MS culture medium, in different concentrations of growth regulators, including four levels of TDZ (0.5, 1, 2, and 3mg/L), two levels of IAA (0.5 and 1mg/L), and four levels of adenine hemi-sulfate (0. 20, 40, and 80mg/l), were used.

The model used for this experiment was significant at the level of 1%, where R2=96% and CV=22, indicating the acceptable accuracy of the analysis. With respect to the results, it was observed that, in the leaf explants of different carnation cultivars, the utilization of the MS culture medium in light conditions with TDZ 2 mg/L+ IAA 0.5 mg/L occurred the direct regeneration of the leaf explants. It is worth mentioning that the addition of 40mg/L of adenine hemi-sulfate to the culture medium increased the regeneration mean from 66% to 94% among different cultivars. In addition, the mean shoot number per explant increased from 12 to 36 shoots.

Generally, the results of the experiments showed that the mixing of the MS base medium, leaf explants in light conditions, and the TDZ 2 mg/L+ IAA 0.5 mg/L+As 40 mg/L regulator composition significantly led to the leaf regeneration of carnation cultivars. Briefly, the results of this study proved that the use of adenine hemi-sulfate significantly impacted the upsurge of the direct regeneration rate of carnation cultivars.

To optimize tomato (Solanum Lycopersicum) plant tissue culture condition, the effect of 13 different hormonal combinations and two explants of cotyledon and hypocotyl on the regeneration rate of three Newton, Infinity and Defines cultivars were investigated. Experiments were conducted in a factorial based on a completely randomized design with three replications. The highest shoot number (11.9 shoots per explant) was obtained for Defines cultivar on MS medium containing the hormones of TDZ=0.68, NAA=0.1milligrams per liter and cotyledon explant. Plant defense responses against environmental factors are one of the inhibitor factors of explants regeneration in plant tissue culture. Calcium ion signaling is one of the primary events and, of course, the most important event in plant defense response against pathogens, Therefore, the effect of verapamil as a calcium channel blocker was investigated on the plant defense response. For this purpose, inoculation was performed using Agrobacterium tumefaciens strain LBA4404 and based on optimized tissue culture condition. The effect of verapamil in four concentrations of 0, 50, 100 and 200 milligrams per liter was investigated in a co-culture medium. Results showed that the use of verapamil in comparison with the control sample reduced the plant defense response. Also, the use of 50 milligrams per liter of verapamil increased the amount of regeneration.

Developing a highly efficient protocol for plant regeneration is a prerequisite for successful gene transformation in plants. To gain these properties, several experiments were conducted on various explants to optimize the concentration of plant growth regulators in regeneration of Gerbera. In the first experiment, regeneration of Gerbera plant was investigated using different explants including intact leaf, petiole, scratched petiole and Thin Cell Layer (TCL). Moreover, combination of BAP (0, 1, 2 and 3 mg L-1), TDZ (0, 0.1, 0.5, 0.8 and 1 mg L-1) and IAA (0.1 mg L-1) under 16/8 h light/darkness condition or one-month darkness were considered. In the second experiment, the regeneration potential of the first, second and third disc of TCL cut off from the end of the detached petioles were evaluated. Effect of different combination of BAP (0, 1 and 2 mg L-1), TDZ (0 and 0.5 mg L-1) and IAA (0, 0.1 and 0.3 mg L-1) on regeneration was studied. In all experiments, the MS-basal medium was used. The results showed that the best regeneration medium for intact leaf (33.2% regeneration, 66.3 shoots), petiole (86.66% regeneration, 33.33 shoots) and scratched petiole (53.2% regeneration, 33.10 shoots) was MS medium containing 2 mg L-1 BAP under 16/8 h light/darkness condition, and for TCL explants (46% regeneration, 29 shoots), 2 mg L-1 BAP with 0.5 mg L-1 TDZ in the darkness. In the second experiment, the highest rate of regeneration was obtained from the first disc of TCL explants on the MS medium containing 2 mg L-1 BAP, 0.5 mg L-1 TDZ and 0.1 mg L-1 IAA.

Calla lily (Zantedeschia spp.) is one of the monocotyledonous ornamental plants that is famous as pot plant and cut flower in the world. In the present study, in order to investigate indirect regeneration, different explants of calla lily cv. Orania were used in a Completely Randomized Design (CRD). In order to produce callus, explants of tuber buds, meristem, shoot-tip from tissue cultured plantlets and thin cell layer of stem were cultured on MS medium with different concentrations of BAP and NAA as treatment combination. Afterwards, the effect of using amino acid L-glutamine and adenine sulfate on increasing the shoot induction percentage was investigated. The highest percentage of callus formation (73.3%) was obtained from meristem explant on the MS medium containing 1 mg L-1 BAP along with 1.5 mg L-1 NAA. Due to the good quality of calli induced from the meristem explants, this explant was transferred to MS medium containing BAP, adenine sulfate and L-glutamine for regeneration. Highest regeneration percentage (78.3%), highest average number of shoots (13.4) and average shoot length (51.6 mm) were obtained on the media containing 1 mg L-1 BAP with 20 mg L-1 adenine sulfate and 300 mg L-1 L-Glutamine. Considering that calla lily is a recalcitrant plant in tissue culture, adding 500 mg L-1 activated charcoal to the rooting medium containing 0.25 and 0.5 mg L-1 IAA increased both rooting percentage and number of roots per plantlet. Furthermore, using amino acid L-glutamine (300 mg L-1) and adenine sulfate (20 mg L-1) increased the quality and number of plantlets produced from regeneration. Regenerated plantlets with proper length were transferred to the MS medium containing 0.5 mg L-1 IAA with 500 mg L-1 activated charcoal as rooting medium with the highest percentage of root induction (88.3%). Rooted plants were adapted (more than 70%) successfully in the perlite and cocopeat mixture (1:1) under greenhouse condition.

Gladiolus is cultivated as a high-value ornamental plant in the worldwide and it has the highest cultivation area and the highest level of underground organs imports in Iran. In this study, in order to investigate indirect regeneration of Amsterdam and Advance Red cultivars an experiment was conducted in a completely randomized design with three replications. For callogenesis, bud sprout (apical section), corm slices (middle section) and basal plate (bottom section) explants of corm with 1 cm2 dimensions were cultured in MS medium with different concentrations of Picloram, Dicamba, BAP , kinetin, NAA and 2,4,5-T. At the regeneration stage, different concentrations of TDZ and GA3 were combined with Proline and Glutamine amino acids. Based on the results, in the medium containing 1 or 2 mg L-1 of Dicamba, the bud sprout explant showed the highest callogenesis (77.77% and 75.91% in Amsterdam and Advance Red, respectively). The highest percentage of regeneration (81.11% and 75.57% in Amsterdam and Advance Red, respectively) and the highest number of shoots (23.69 and 20.19 in Amsterdam and Advance Red, respectively) were obtained in 1 mg L-1 TDZ + 0.5 mg L-1 GA3 + 200 mg L-1 Proline + 200 mg L-1 Glutamine. The highest percentage of rooting (84.41% and 82.50% in Amsterdam and Advance Red, respectively) were produced in MS medium supplemented with 2 mg L-1 IAA + 500 mg L-1 activated charcoal. Produced plantlets from indirect regeneration were successfully adapted in greenhouse condition.

In vitro organogenesis of cucumber (Cucumis sativus L.) was evaluated using the cotyledonary explants of “Beth Alpha” and “Dastgerdi” cultivars. Organogenesis was examined on the MS medium containing 2 mgl-1 Benzylaminopurine (BAP) along with eight combinations of abscisic acid (ABA) (0 and 1 mgl-1), silver nitrate (0 and 5 mgl-1), and cefotaxime (0 and 200 mgl-1). The results indicated the significant effect of ABA on increasing the percent of shoot induction and number of regenerated shoots. In the absence of ABA, silver nitrate had a significant effect on shoot induction and number of shoots. Not only the negative effects of cefotaxime was not observed, but also the number of shoots increased in one case. Root induction in the regenerated shoots was also studied using 5 different media including PGR-free MS medium and the media containing indolebutyric acid (IBA) (0.1 and 1 mgl-1) or naphthaleneacetic acid (NAA) (0.1 and 1 mgl-1). The highest number of roots per shoot in “Beth Alpha” (9.4) and “Dastgerdi” (8.6) were observed in the MS medium supplemented with 0.1 mgl-1 IBA. The genetic stability among 10 randomly selected clones of “Beth Alpha” cultivar was evaluated using AFLP markers. Besides that, cytogenetic assessment of 10 clones from each cultivar was implemented. In total 293 monomorphic bands were amplified using 9 primer combinations of AFLP marker, indicating the absence of somaclonal variation. The chromosome counting of root meristematic cells revealed the stability of ploidy level among regenerated clones.

Healthy lifestyle on dry land depends on monitoring and maintenance of the ponds. The aim of this study was to assess the effects of surface water-inflows on biological characteristics of wetland soils around Arak Meyghan-Lake. Two soil layers (0-30 and 30-60 cm) in the 4 sites of surface water-inflows were sampled in May and November 2014. The sites were the release sites of treated-municipal-wastewater, Shahrab-River, Farah-River and sodium sulfate-processing plant wastewater. The number of culturable microorganisms in the samples was counted on solid media and the soil basal-respiration (BR), substrate-induced-respiration (SIR), metabolic-quotient (MQ) and microbial-coefficient (MC) were measured. The number of soil fungi in the site Farahan-River site and the numbers of soil bacteria, enteric bacteria and actinomycetes in the release site of municipal wastewater especially in topsoil and in spring were significantly higher compared to those in the 3 other sites (respectively 4.85, 6, 5.51 and 3.92 log numbers). The numbers of the studied microorganisms in soil in the release site of sulfate-plant wastewater were significantly low and the numbers of fungi and actinomycetes were negligible in the most samples. Soil SIR was the most susceptible and responsive index among the studied biological properties. The release of municipal-wastewater in the playa improved the soil biological indices like BR (0.102 mg CO2 g-1 soil day-1), SIR (3.688 mg CO2 g-1 soil day-1), and MC (0.196 mg Cmic mg-1 Corg) significantly. In contrast, the MQ was significantly high in the release site of Shahrab-River and sulfate-plant wastewater (2.60 and 2.52 (mg C-CO2 g-1 Cmic h-1) respectively), showing the negative effects and the higher salt tension in these sites. Although the release of municipal-wastewater in Meyghan-Lake had positive effects on soil biological characteristics, it may have negative effects on biodiversity of these salty soils.

Greater celandine (Chelidonium sp) is one of the plants that its propagation through seed occurs slowly. In addition, Chelidonium majus L. has limited habitats in Iran. For this reason, micropropagation can be considered as an effective method for its rapid and massive propagation and conservation, which can lead to the production of highly uniform plants. Chelidonium majus L. also contains a large amount of secondary metabolites of Isoquinoline alkaloids, including Chelidonine, Sanguinarine, Captesin, Berberrine and Chloritrine, and phenolic compounds. Therefore, the aim of this study was to investigate micropropagation of Chelidonium majus L. and compare the total phenol content in leaf, stem and root in obtained plantlets.

To begin the experiment, seeds of Chelidonium majus L. were first washed with distilled water containing a few drops of tween20. Then they were washed with 70% alcohol for 1 min and were finally washed with double-distilled water. Next, they were disinfected with 1% sodium hypochlorite for 5 min, and again were rinsed with distilled water for 3 times of 5, 15, and 180 min under laminar air flow hood. The effects of TDZ at concentrations 0, 0.25, 0.5, 0.75 and 1 mg/L, BAP at concentrations 0, 0.5, 1, 1.5, 2 mg/L considered. Then, the effect of best treatment in combination with NAA and IBA at 0.25, 0.5, 0.75 and 1 mg/L on growth parameters (number of shoot, shoot length and shoot formation capacity index), were studied. The effect of IBA, NAA and IAA at 0, 0.5, 1, 1.5 and 2 mg/L on rooting parameters (number of root and root length) in MS medium supplemented with 3 g/L activated charcoal in in vitro conditions were evaluated. Then different ratios of cocopeat, perlite and peat moss were used for acclimatization of the obtaining plants. Folin method was used to measure total phenol content. The experiment was conducted as factorial in a completely randomized design with four replications.

The results of analysis of variance for proliferation and rooting traits showed that there were significant differences among the treatments at 1% probability level. The results of means comparison showed that the highest numbers of shoots and shoot formation capacity index were obtained  from the treatment of 0.5 mg/L TDZ with the average of 8.12, which did not show a significant difference from the concentration of 0.25 mg/L TDZ, and the lowest shoot number was related to the control treatment. Increasing the amount of TDZ hormone led to the reduction in shoot number, so that at concentration of 1 mg/L TDZ, the average shoot number per explant was four. Combination of 0.5 mg/L TDZ with IBA and NAA had lower effect on Chelidonium majus L. proliferation. Moreover, the greatest shoot length was observed in the treatment of 2 mg/L BAP. Comparison of means values showed no significant difference between the treatments of 2 and 1.5 mg/L BAP at 1% probability level. In this study, MS medium containing 1.5 mg/L IBA was the most appropriate treatment for root formation. The effect of NAA hormone on root number of Chelidonium majus L. showed that the highest number of root was obtained from the treatment of 2 mg/L NAA. Besides, the effect of IAA on root number of Chelidonium majus L. showed that the highest number of root was observed in the treatment of 1 mg/L IAA, and the lowest number of root was related to the control treatment .The results of means comparison for the percentage of acclimatized plants showed that the ratio of 0:2:1 had a significant difference from the rest of the culture media and 85% of the plants were acclimatized, while the ratio of 1:2:1 showed the lowest percentage of acclimatization (20%). Furthermore, the results showed that the culture media had significant effect on acclimatization stage at 1% probability level. The results of the analysis of variance for total phenol content in leaf, stem and root tissues showed that there were significant differences among these three tissues. The results showed that the amount of total phenol in leaf was higher than in the stem, and the amount of phenol in root was insignificant.

Based on the results of this study, micropropagation can be used as a method for commercial production of this species under in vitro conditions.

Due to the lack of water in different regions of Iran, the introduction and use of indigenous species with valuable ornamental features such as mullein (Verbascum thapsus) is very important. V. thapsus is a perennial and evergreen herb belonging to the Scrophulariaceae family, which like other ornamental plants, can be affected by drought stress in the green space. Drought is an environmental stress that induces adverse effects on most stages of growth, structure and activities of plants. The response of plants to environmental stresses is different in morphological, cellular and molecular levels. Therefore, the present study was conducted to investigate the effect of drought stress on morphological, physiological and biochemical characteristics of V. thapsus during two stages of vegetative and reproductive growth.

In the present study, the reaction of V. thapsus to drought stress and its effect on vegetative and reproductive stages were investigated. The experiment was carried out as factorial in a completely randomized design with two factors as drought stress in 5 levels (control (0.3), 2, 5, 10 and 15 bar) and plant growth stages at two levels (vegetative and reproductive). For this purpose, shoot and root fresh weight, burn and wilt condition, leaf chlorophyll, activity of antioxidant enzymes and proline were measured.

The highest and lowest irrigation interval in both vegetative and reproductive stages was respectively found in 15 bar and control. The water volume used in vegetative stage was higher than reproductive stage. Shoot and root fresh weight in was reproductive stage was more than vegetative stage. The highest fresh weight of shoots was reported under drought stress in control and 2 bar and the lowest amount was observed in 15 bar, but the highest root fresh weight was observed in 5 bar. The highest percentage of leaf burns and wilt was observed in treatment of 15 bar × reproductive stage and its lowest in the treatments of control × vegetative stage and control × reproductive stage. Total chlorophyll in reproductive stage was more than vegetative stage. Also, the highest total chlorophyll was observed under drought stress in control and 2 bar and the lowest was found in 15 bar. The highest activity of superoxide dismutase and catalase was observed in 15 bar × reproductive stage and its lowest value was recorded in control × vegetative stage. Proline in reproductive stage was greater than the vegetative stage. The highest and lowest amount of peoline was observed in 15 bar and control, respectively.

The results of the research showed that the drought stress up to 5 bar did not cause significant changes in the plant, but increasing the intensity of drought stress from 5 bar to 10 bar induces the significant change of most traits in the plant. Therefore, with applying an appropriate plan, we can reduce the water use from 570 m3 (control) to 130 m3 (5 bar) during the vegetative stage and 500 m3 to 140 m3 for reproductive stage.

Benefits of cut style method and ovary slice culture for overcoming pre-and post-zygotic barriers in lily crossing were studied. Three Asiatic hybrid cultivars (Tressor, Pirandello and Ceb duzzel) along with four cultivars of Oriental section (Rialto, Sorbonne, Paradero and Manissa) were used as maternal plants. Lilium ledebourii has selected as pollen donor plant to cross with maternal plants using stigma pollination or cut style pollination methods. Harvesting of produced capsules for ovary slice culture was done after 10, 20 and 40 days. Ovary expansion index, number of produced capsules, capsule weight, and number of cultured slices, number of produced seeds, seed weight and number of germinated seeds were recorded. Results indicated that cultivar had significant influence on all recorded parameters. The number of seeds was significantly affected by capsule harvesting time (p<0.05). In cut style experiment in crosses between Lilium ledebourii and Mannisa, Tressor and Sorbonne 172, 80 and 60 seeds were produced respectively. The rate of seed germination for Mannisa by L. ledebourii was 1.1 percent. In stigma pollination experiment capsules were expanded but none of them produced viable seeds. Totally, in this experiment, use of CSM pollination method was successful for by passing pre-zygotic barriers in lily interspecific crossing.

Chrysanthemum is one of the ornamental plants that cultivated widely in worldwide. This flower is one of the most important cut and pot flowers in the world due to its high diversity of colors and shapes. The creation of mutation is an important method for the production of new cultivars, and many cultivars have been produced through spontaneous and induced mutations. In this study, explants of leaf pieces from three most cultivated chrysanthemum varieties were irradiated with different gamma rays. This experiment was conducted as factorial in randomized complete block design with three replications. The results showed that an appropriate dose of gamma ray to produce a mutation in the varieties used in this experiment, is 25 G-ray. The obtained results showed that the 25 G-ray in purple cultivar produced the greatest change in petal color with a mutation rate of 54.56%. Meanwhile, the maximum number of new colors belonged to the purple group. Also, in the pink cultivar, the highest number of colored flowers was observed with a change of 32.11% in the 25 G-ray treatment. Based on the results of this study, four new cultivars will be introduced to the Iranian flower industry as a new cultivar.

Gerbera is one of the most popular ornamental plants in the world and ranked forth, in the flower industry among the 10 top cut flowers. Nowadays, there is an important for development of generally applicable gene transfer methods for gerbera which will allow rapid introduction of new traits into elite genotypes without changing the existing good properties. This study was conducted in three separate experiments; First experiment was conducted to evaluate the effect of hgygromaycin at various concentrations (5, 10, 15, 20, 25 and 30 mg L-1) to determine it’s the lethal dose in untransformed plantlets of Gerbera jamesonii cv. ‘Royal Soft Pink’. Then, in second experiment investigated the effect of 10, 15, 20, 25 and 30 mg L-1 hgygromaycin on shoot regeneration in leafy petiole explants. Finally, in third experiment evaluated the effect of inoculation times (10 and 20 min) and acetosyringone concentrations (50 and 100 µmol) on GUS gene transfer efficiency mediated by Agrobacterium and by co-cultivation method of leafy petiole explants. After 2-3 days of co-cultivation of leafy petiole explants with Agrobacterium, were transferred to direct shoot regeneration medium. Then putative transgenic shoots derived from petioles and the multiplication were done. Finally, transformed plants were confirmed by GUS histochemical assay and PCR analysis. The results showed that the 10 mg L-1 concentration of hgygromaycin is suitable for selection of transgenic shoots and it should be used after initial regeneration. The highest (38%) of putative transgenic shoots regeneration was obtained from treatment of 10 min inoculation with Agrobacterium and using 100 µmol acetosyringone. Also, the lowest (25.25%) of putative transgenic shoots regeneration was showed from treatments of 20 min inoculation and using 50 µmol acetosyringone. The results of histochemical assay and PCR analysis in transgenic plants showed that the highest independent line number (27 shoots) and gene transfer efficiency (11%) per 100 explants were obtained from treatments of 10 min inoculation and using 100 µmol acetosyringone. Factors such as inoculation time and acetosyringone concentration had prominent impact on the efficiency and success of transformation. Acetosyringone, is considered to be the phenolic inducer of vir genes activation and T-DNA transfer in Agrobacterium and enhances the transgenic efficiency. Therefore, the best treatment to produce transgenic plant in Gerbera jamesonii cv. ‘Royal Soft Pink’ with transfer of GUS gene is the inoculation of leafy petiole explants for 10 min and using of acetosyringone with 100 µmol concentration

Effect of different temperature treatments on the biochemical compounds and growth parameters of Hyacinthus orientalis cv. Blue jacket bulbs were evaluated under different temperature (5, 10, 15 and 20 °C). After four weeks, biochemical traits such as carbohydrate solution, protein, triglyceride, peroxidase and amylase during storage were measured. After the storage period they were planted in pots and some physiological traits during season growth were examined. Storing at 5 and 10 °C showed the highest carbohydrate solution, protein and triglyceride content and highest enzyme activities were observed in 15 and 20 °C treatments. Moreover, during the growing season the plant treated with 5 and 10 °C showed higher growth rate and earlier flowering time. Overall, according to the results, to achieve high quality and rapid flowering, storage temperature of 5 or 10 °C are recommended in hyacinth bulbs before planting.

Saffron (Crocus sativus L.) is the world's most expensive spice. Moreover, it is important since it contains various drug metabolites. Saffron is a triploid (2n=3X=24) and sterile plant and it does not have any viable seeds. Because of the sterility, classical breeding of this plant is limited. Developing an efficient callus induction protocol is studied for two reasons, i.e. molecular breeding and the production of secondary metabolites. In order to provide a suitable callus induction protocol, establishment of corms was considered. Five different treatments were applied to sterilization of corms. Thin cell layer explants with approximately 1 mm thickness and typical explants with approximately 1 cm thickness were prepared from sterilized corms. For callus induction, different explants were planted in MS medium containing different concentration of 2, 4-D, BAP and NAA. Then, they were incubated in dark conditions at 20 ±20C for 3 months. The results showed that the use of Benomyl fungicide, followed by surface sterilization using sodium hypochlorite (2.5 %) was the best sterilization treatment. The highest survival rates of explants (90%) were observed in this treatment and all explants were free of contamination. The highest amount of callus induction (75%) was obtained in MS medium supplemented with 2mg/l NAA and 0/5 mg/l BAP from thin cell layer of basal corm. The same result was observed with 1mg/l 2, 4-D from typical explants of basal corm. The results obtained from this study show that the thin cell layer explants are suitable explants because of the high amount of callus formation and the advantages for gene transfer studies.
The results showed that the use of Benomyl fungicide, followed by surface sterilization using sodium hypochlorite (2.5 %) was the best sterilization treatment. The highest survival rates of explants (90%) was observed in this treatment and all explants were free of contamination. The highest amount of callus induction (75%) was obtained in MS medium supplemented with 2mg/l NAA and 0/5 mg/l BAP from thin cell layer of basal corm. The same result was observed with 1mg/l 2,4-D from typical explants of basal corm. Our results showed that the thin cell layer explants are suitable explant because of high amount of callus formation and advantages for gene transfer studies.

The study of flower color genes function in gerbera is hampered due to the low efficiency of transformation methods and the long time span needed for production of stably transformed transgenic plants (1). For some functional analysis, the transient expression of genes could be an efficient alternative.
Therefore, this study was conducted in two stages. In first stage, the agroinfiltration experiment with 3 flower color constructs (1-pBIH-35S-CcF3´5´H: with one gene, 2-pBIH-35S-Del2: with 3 genes and 3-pBIH-35S-Del8: with 5 genes) in 12 cultivars of gerbera was investigated. Agroinfiltration of gerbera petals were performed by Agrobacterium tumefaciens strain EHA 101 harboring binary vectors pBIH that contained one or more genes of flavonoid 3ʹ 5ʹ-hydroxylase (F3´5´H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), flavanone 3β-hydroxylase (F3H), chalcone isomerase (CHI) and hygromycin phosphotransferase (hpt).
Visual observations of injected petals showed that cultivars with pink color have shifted flower color from pink to blue and produced delphinidin. In the second stage, this experiment was repeated with 4 pink cultivars (‘Aqua Melone’, ‘Bismarck’, ‘Esmara’ and ‘Rosalin’) and mentioned constructs. The results of HPLC analysis of 4 anthocyanins (delphinidin, cyaniding, pelargonidin and peonidin) showed that the injected petals of ‘Bismarck’ cultivar with pBIH-35S-Del8 construct have the highest delphinidin production.
Therefore, we can suggest ‘Bismarck’ cultivar of gerbera for stable transformation of genes involving production of anthocyanins for change of flower color particularly production of delphinidin.

The genus of Hawthorn (Crataegus L.), belonged to Rosaceae family has medicinal and ornamental applications. Its seed germination is poor, due to stony endocarp and embryo dormancy. This study was carried out to determine methods to break dormancy in Crataegus pseudoheterophylla seeds. For this purpose, seeds were sown in plastic pots as a factorial experiment in randomized completely block design (RCBD) with 3 replications. Treatments including chemical scarification in KNO3 for 0, 2500, 5000, 7500 and 10000 mg l-1 and concentrate sulfuric acid (H2SO4 96 %) for 0, 30, 45, 60, 90, 120, 150, 180 and 210 minutes followed by continues regime (1 month at 23 °C, then 3 months at 4 °C) and alternate regime (1 month at 4 °C and, 2 weeks at 23 °C, then 1 month at 4 °C and 2 weeks at 23 °C, finally 1 month at 4 °C) were applied. Results showed that in KNO3, the highest germination percentage (19.33 %) and germination speed (4.18 number/day) was determined at 10000 mg l-1 alternate stratification and in sulphuric acid, the highest percentage germination (55 %) and speed germination (8.63 n/d) was determined for 120 minutes alternate stratification. The highest mean time germination in both scarification treatments was observed in continues regime.

RNA extraction from hard wood tissues, especially in coniferous family trees, is very hard because they have high concentrations of phenolic compounds, polysaccharides, secondary metabolites and lignin. Moreover, up to now, no commercial kit is made for the mentioned purpose. In this study, the problems were resolved and a rapid, simple, and economic protocol is presented for RNA extraction from bark tissue of Cedrus deodara and Abies grandis, which belong to conifer family. Ratios of optical density of 260/280 nm and 260/230 nm were respectively obtained as 1.88 and 1.1 for Cedrus deodara and 1.8 and 0.9 for Abies grandis. Modifications for optimization of RNA extraction were preheating treatment of extraction buffer, and decreasing centrifuge rounds to eliminate DNA contaminations and DEPC. Synthesis of cDNA was performed using extracted RNA. RT-PCR successfully produced expected bands. Therefore, it seems that the presented protocol is suitable for RNA extraction from woody tissues.