Breeding and Biotechnology of Plant and Flower
Letafat Moradi; Seyyed Najmodin Mortazavi; Mitra Alaei
Abstract
IntroductionSuccess in tissue culture technique, especially in bulbous plants, depends on the microbial contamination control during in vitro culture. Decontamination is considered as a fundamental challenge in the technique of cell, tissue and plant organ culture. Although there are various methods ...
Read More
IntroductionSuccess in tissue culture technique, especially in bulbous plants, depends on the microbial contamination control during in vitro culture. Decontamination is considered as a fundamental challenge in the technique of cell, tissue and plant organ culture. Although there are various methods for this purpose, the development of disinfection methods specific to each species is considered an important factor in the establishment and success of the tissue culture system. Applying different treatments can control the microbial contamination and consequently increase the percentage of explant survival. This plant is among the poisonous plants of pastures and the chemical compounds present in it have important medicinal effects, so that the existence of alkaloid and glycosidic compounds has been reported for it. These chemical compounds are used to treat rheumatic pains, lymphatic system diseases and also as liver cleansers. This ornamental plant also has the ability to produce pots and its export is possible in pots . Currently, in order to overcome the issues of the usual methods of propagation of this ornamental plant and to produce pollution-free plants, the use of in vitro cultivation methods in this plant has become very important. bulbes is one of the most common explants of inverted tulip tissue culture, but it often faces very high internal and external fungal and bacterial contamination. In the in vitro culture of onion plants, the existence of fungal and bacterial contaminations are one of the main problems that can affect the efficiency of this type of propagation methods. Fungal and bacterial contaminations are one of the most important problematic factors in different in vitro culture methods. Internal contamination usually appears several weeks after planting. These contaminations may not be seen externally, but they affect the growth, division and greening of small samples. Mercuric chloride is mainly used as a surface disinfectant along with sodium hypochlorite and it controls the spectrum of bacteria and fungi and increases the disinfection efficiency. The results of the study showed that mercuric chloride relatively controls the bacterial and fungal contamination of the samples, but it reduces the survival percentage of the samples in high concentration. It seems that some plants are sensitive to mercuric chloride and determining the appropriate amount of this substance plays a very important role in preparing a healthy and living specimen. Determining the duration of disinfectant treatment was also considered in various researches, so that sometimes carbendazim 1% fungicide for 20 minutes, 70% alcohol for 60 seconds, sodium hypochlorite 2.5% for 10 minutes. And 0.1% mercury chloride was used for 7 minutes to sterilize aloe vera sample. Considering the importance of plant tissue culture techniques and the need to create new protocols to reduce contamination and increase the number of healthy cultures, the main goal of this study is to create an efficient protocol for the disinfection of Fritillaria spp. onion explants in order to increase the efficiency of in vitro cultures. Materials and Methods This study aimed to investigate the effect of time and different concentrations of sodium hypochlorite, carbendazim fungicide and mercuric chloride in inhibiting the contamination of inverted tulip bulbs (Fritillaria spp.) in tissue culture environment. So, an experiment was done as a completely randomized design at four replications in the biotechnology laboratory of Zanjan University during 2023. The experimental treatments consisted of 0.1% fungicide at different times (30, 25, 35 and 40 minutes), 5 levels of sodium hypochlorite (0,1, 1.5, 2, 2.5 and 3%) at different times (7, 9, 10, 12 and 15 minutes), 70% Ethanol at two different times (0,60 and 90 seconds) and mercury chloride (0,0.1 and 0.2%). Bulbs that collected from nature were transferred to the Biotechnology Laboratory of the Faculty of Agriculture, Department of Horticulture, Zanjan University and kept them in the refrigerator at 4°C for two weeks. After this period of time, the bulbs were washed with detergent and then remained in water for 30 minutes. Then they were disinfected by using the above-mentioned treatments: It should be noted that at the end of each step, Fritillaria bulbs were washed with sterile distilled water. Also, after the end of the disinfection treatments, the desired explants were cultured in MS basic culture medium. The statistical analysis of this experiment was carried out using SAS software, version 9.1. The differences between mean values were compared using Duncan’s multiple range test method at the 5% significant level (p < 0.05). Results and Discussion The control of fungal and bacterial contamination, especially in relation to bulbous and bulbous plants that are in direct contact with the culture medium, is one of the basic steps in in vitro culture conditions. Applying different treatments including the use of fungicides, alcohol, sodium hypochlorite and mercuric chloride in the cultivation environment can help control fungal and bacterial contamination in the Fritillaria spp. plant. However, the application of such treatments can have a negative effect on the regeneration potential of the explants, so it is important to choose the appropriate treatment in a way that controls the fungal and bacterial contamination and on the other hand preserves the regeneration potential of the explants. This study investigated the effectiveness of various disinfection protocols on Laleh-Avgagun onion explants. While the lowest contamination (0%) was achieved using 0.1% fungicide for 40 minutes, 70% alcohol for 90 seconds, 3% sodium hypochlorite for 15 minutes, and 0.1% mercury chloride, this treatment significantly reduced explant survival and regeneration, causing browning. Conversely, applying 0.1% fungicide for 30 minutes, 70% alcohol for 60 seconds, 2% or 2.5% sodium hypochlorite for 12 or 10 minutes respectively, and 0.1% mercury chloride effectively reduced contamination while maintaining explant survival. This optimized protocol improved the growth and regeneration ability of the explants.Therefore, in the conditions of in vitro cultivation, the use of the mentioned treatments is recommended to control contamination and optimal reproduction of the bulbs of Fritillaria spp. plant.
Mina Ghazaeian; Gholamhossein Davarynejad; Kamal Ghasemi Bezdi; Seyyed Hossein Nemati
Abstract
Introduction: 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 ...
Read More
Introduction: 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.
Materials and Methods: 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.
Results and Discussion: 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.
Conclusion: 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.
Masud Ghasemi; Hossein Arouiee; Pejman Azadi; Atoosa Ali Ahmadi
Abstract
Introduction: 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, ...
Read More
Introduction: 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.
Materials and Methods: 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.
Results: 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.
Conclusion: Based on the results of this study, micropropagation can be used as a method for commercial production of this species under in vitro conditions.
Ahmad Sharifi; Azadeh Khadem; Maryam Moradian; Seyyedeh Mahdiyeh Kharrazi
Abstract
Introduction: Application of stress-tolerant rootstocks is known as one of the effective methods to enhance the productivity of fruit gardens. Pear is one of the four or five major classes of fruits that are produced worldwide. Among pear cultivars, Pyrodwarf rootstock is tolerant to alkaline soil and ...
Read More
Introduction: Application of stress-tolerant rootstocks is known as one of the effective methods to enhance the productivity of fruit gardens. Pear is one of the four or five major classes of fruits that are produced worldwide. Among pear cultivars, Pyrodwarf rootstock is tolerant to alkaline soil and low temperatures of winter. In addition, many cultivars of pear with high productivity could be grafted to this rootstock. These advantages result in widespread culture of pyrodwarf rootstock in pear gardens. The increasing market demand of this rootstock presents an opportunity to investigate alternative methods for efficient production of pyrodwarf. Therefore, this study has been conducted to increase the efficiency of pyrodwarf propagation through tissue culture methods.
Materials and Methods: For this experiment, sterile plantlets of pyrodwarf rootstocks in a same growth phase were used. Plantlets were cultured in different 12 media including MS, WPM, KNOP, modified KNOP, QL and modified QL basal media supplemented with 0.5 mg.L-1 BA and 0.05 mg.L-1 IAA or IBA and solidified with 8 g.L-1 agar. After that, cultures were transferred to the growth room at 25 ± 1°C under a 16/8h light/dark photoperiod (light intensity 30 μmol.m−2.s−1, cool-white fluorescent light). In the next step for rooting optimization, two separate experiments were conducted. In the first, 10 different basal media containing various nutritional compounds were examined for rooting properties of pyrodwarf plantlets. The following experiment was performed to reveal the effects of IAA and active charcoal on the efficiency of plantlets rooting. All the experiments were carried out in acompletelyrandomized design. The study of propagation efficiency of plantlets was arranged in a factorial approach with twofactors including the type of basal media (sixlevels) and auxins (twolevels) with threereplications. The both experiments of plantlet rooting were performed with sevenreplications. Data were analyzed using JMP-8 statistical software and the meanswere compared by using LSD test at 5% probability.
Results and Discussion: All explants were regenerated 15 days after cultivation. The number of buds per explants indicates that nutritional compounds significantly affected bud formation. MS basal media induced the most number of adventitious buds and decreasing amount of nitrogen in the culture medium (either in its ammonium or nitrate form) leads to 50%-decrease in the number of buds induced per explant. Previous reports mentioned that macronutrients have the most significant effect on bud formation of peach hybrid rootstock GF677. In addition, high amount of nitrogen is necessary to achieve the highest height of pyrodwarfin vitro plantlets. In contrast to the basal media, neither bud formation nor height of plantlets did not affected by the type of auxins, IAA and IBA. Also weight of plantlets was similar in all treatments and shows the effect of basal media and auxins were not significant on biomass production. According to the results of rooting experiment, plantlets that were cultured in MS and DKW had higher height than QL and Knop in rooting phase. Also, 1/2 MS-cultured plantlets have the lowest height among all treatments. These results showed that macronutrients have positive effects on the growth of pyrodwarf plantlets as decreasing concentration; the growth of plantlets was inhibited. In contrast to shoot growth, number of roots per explants in 1/2 MS medium was significantly higher than the other media. These results are expected as shoots and roots of plants have different response to environmental stimuli. Data obtained from the following experiment of rooting indicated that IAA improved root induction similar toother functional traits of plants but the presence of both of those compounds in medium have inhibitory effect on the functional traits of plants.
Conclusion: These data suggest that macronutrients and hormonal compounds have impressive impact on the regeneration of pyrodwarf plantlets. In this regard, MS medium supplemented with 0.5 mg.L-1 BA and 0.05 mg.L-1 IAA or IBA induced most number of buds and the regenerated plantlets hadthe highest height in that medium compared to the other treatments. Also, 1/2 MS medium containing 1 mg.L-1 IAA was identified as the best media in case of rooting parameters.
Ahmad Sharifi; Fatemeh Keykha Akhar; Mahboobeh Yazdi; Abdolreza Bagheri
Abstract
Introduction: 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 ...
Read More
Introduction: 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.
Materials and Methods: 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).
Results and Discussion: 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.
Asad Masoumiasl; Ameneh Aryiaeineghad; Masud Dehdeari
Abstract
Introduction: German (Matricaria chamomilla L.) and Shirazi (Matricaria recutita L.) chamomiles are the most important medicinal plants of the Astraceae family which are used in the pharmaceutical, health, food and cosmetics industries. Production of this plant has been undertaken in Iran mainly in Isfahan, ...
Read More
Introduction: German (Matricaria chamomilla L.) and Shirazi (Matricaria recutita L.) chamomiles are the most important medicinal plants of the Astraceae family which are used in the pharmaceutical, health, food and cosmetics industries. Production of this plant has been undertaken in Iran mainly in Isfahan, Kohgiluyeh and Boyerahmad, Golestan and Hamedan provinces. In vitro propagation of plants have higher potential to produce qualified natural products, restoring and preserving of endangered plants, induction of somaclonal variation, industrial reproduction, valuable secondary metabolites and increased active ingredients. Researchers reported successful micropropagation system for five chamomile varieties on MS medium contained 0.01 mg/l NAA and 2.5 mg/l kinetin. The aim of this study was to investigate the effects of stem (with and without node), leaf and cotyledon explants and different plant growth regulators on direct regeneration of German and Shirazi chamomiles. Although the Shirazi chamomile is native to Iran, but German chamomile is Iran non-indigenous cultivar. By our knowledge, there have been no comparison reports about responses of these cultivars to tissue culture.
Materials and Methods: This research was performed in the central laboratory of Agriculture Faculty in Yasouj University. Seeds were provided from Pakan-Bazr institute, Isfahan. Chamomile seeds were disinfected by ethanol (70%) for 5-10 min and sodium hypocholorite 3% for 5-12 min and then washed for several times by distilled water. Then, seeds were sown on MS medium for germination. After 2-3 weeks, seedlings were grown and then planted in MS medium supplemented with hormonal combinations of NAA at two levels (0.1 and 0.5 mg/l), kinetin at three levels (2, 2.5 and 3 mg/l) and Zeatin, BAP and 2ip at three levels (0.5, 1 and 1.5 mg/l). The experiment was performed in a completely randomized design with four replications. Factors included explants, cultivars and hormonal combinations. In this experiment, traits such as stem induction percentage, stem length, stem fresh and dry weight root induction percent, root length and stem fresh and dry weight were measured. Statistical analysis was performed using SAS software (version 9.1). In order to test the normality and perform mean comparisons, Minitab 14 and MSTAT-C software was used and excel software was used for drawing diagrams.
Results and Discussion: For all traits except stem induction percent and root dry weight, triple interaction of explants, hormonal combinations and cultivar were significant at 1% level. Triple mean comparisons for cultivar, explant and hormonal combinations shows that the highest mean for stem height, stem fresh weight, stem dry weight, root induction percent, root height and root fresh weight was obtained in Shirazi Chamomile cultivar with cotyledon explants in hormonal combinations of 0.1 mg/l NAA and 1.5 mg/l 2ip. The results showed that the best explants in both chamomiles for direct regeneration were stem (whit node) and cotyledon with 78.75% and 75% regenerations, respectively. Also the best genotype and hormone combination were Shirazi chamomile and MS medium supplemented with 1.5mg/l 2ip with 0.1mg/l NAA. To determine the best hormonal combination for root regeneration from direct regeneration, the stems were embedded in medium contained different concentrations of IBA. The first signs of rooting production were observed after 5-7 days. After completing the roots formation (4 weeks after transferring), the rooting percentage, root length, fresh and dry weight of roots were measured. Based upon variance analysis, effect of triple interactions of IBA, explants and cultivar on all traits were not significant, but the effect of IBA for all traits was significant at 1% level. According to the results of mean comparisons for effects of IBA on root traits in direct regeneration of chamomile, the highest percentage of root regeneration (73.75 %), the highest root length (6.60 cm), root fresh weight (174.167 mg) and the highest root dry weight (16.425 mg) were obtained from medium contained 0.5 mg/l IBA. Root differentiation was influenced by auxin (0.5 mg/l IBA) or spontaneously. Regenerated plantlets were transferred to pots contained sterilized soil (3:1:1 mixture of soil: sand: leaf compost). For plant adaptation to natural conditions, glassy caps were used. After adaptation, the caps were removed and the plants were transferred into a growth chamber. Previous studies reported that MS medium supplemented with 0.2 to 1 mg/l of BA and 2 mg/l of NAA induced adventitious bud formation and shoot development in leaf explants of Roman Chamomile. A higher number of adventitious buds were observed at the proximal end of the explants. Plantlets were rooted on MS medium supplemented with 0.1 mg/l of IBA and successfully weaned in vivo.
Conclusion: Based on the results of this research, chamomile showed relevant response to direct regeneration.
E. Chamani; M. Taheri
Abstract
Three experiments were conducted in tissue culture and biotechnology laboratory of Horticulture Department of Mohaghegh Ardabili University in 2012. For the regeneration of plant from seed, different concentrations of NaOH (5, 10, 15, 20 M) and various scarification methods with sandpaper (soft scarification, ...
Read More
Three experiments were conducted in tissue culture and biotechnology laboratory of Horticulture Department of Mohaghegh Ardabili University in 2012. For the regeneration of plant from seed, different concentrations of NaOH (5, 10, 15, 20 M) and various scarification methods with sandpaper (soft scarification, medium scarification and hard scarification) were used based on completely randomized design with 4 replications. The results of experiment revealed that seeds treated by 20 M NaOH and hard scarification produced the highest germination rate. After 2 months of seed germination, hypocotyles of seeds were used as explants and cultured in MS medium containing different concentration of 2,4-D, picloram, TDZ and BA (1, 2, 4 mg/l) based on completely randomized design with 4 replications. Mean comparison revealed that explants treated by 4 mg/l picloram and 1 mg/l 2,4-D produced the highest callus content. Mean comparison showed that explant treated by 1 mg/l BA produced the highest shoots. However, to investigate the soluble protein changes during growth stages and to study the effects of 2,4-D, picloram, TDZ and BA on soluble protein and experiment was conducted. The result showed that by increasing the plant age, soluble protein was reduced and also the highest soluble protein was found after 4 weeks of germination. The result also showed that explants treated by 4 mg/l picloram and 1 mg/l BA produced the highest soluble protein content.
S. Hajian; S. Alizadeh Ajirlo; F. Zare Nahandi
Abstract
Micropropagation is a proper approach to rapid and large-scale propagation of rootstocks and rose cultivars for huge demand of flower market. Proliferation rate of shoot is decreased drastically following several subcultures. Growth regulators have remarkable effects on the key phase of proliferation ...
Read More
Micropropagation is a proper approach to rapid and large-scale propagation of rootstocks and rose cultivars for huge demand of flower market. Proliferation rate of shoot is decreased drastically following several subcultures. Growth regulators have remarkable effects on the key phase of proliferation in micropropagation of this popular crop. In this research the effects of BAP and antiauxin of TIBA on quality and quantity of developed shoots in Rosa hybrida cv. Full House were studied. BAP and TIBA were applied at three concentrations of 0, 2.2 and 8.8 µmol in proliferation phase of micropropagation. The experiment was conducted based on factorial and completely randomized design with four replications. After two months, the percentage of proliferated explants, survived main and lateral shoot number, length of the main and lateral shoots, number of green leaves on the shoots, the average number of shoots with chlorotic and necrotic leaves, the average axillary shoot base diameter, fresh weight of shoots and number of shoots with necrotic tip were recorded. Analysis of variance indicated that BAP was ineffective on the number of the main shoot green leaves and decreasing number of shoots with necrotic tip, but enhanced other traits. The concentration of 8.8 µmol of BAP had greater effect than 2.2 µmol of this growth regulator on mentioned traits. The higher concentration of TIBA resulted to more shoot with necrotic tip. This antiauxin had anegative impact on shoot fresh weight, but the other parameters were not significantly affected.
