Breeding and Biotechnology of Plant and Flower
Ahmad Sharifi; Maryam Moradiyan; Nasim Safari; Azadeh Khadem; Seyyedeh Mahdiyeh Kharrazi
Abstract
Introduction: Ornamental foliage plants are commonly used for beautifying indoor spaces. Consequently, determining the best method of mass propagation in a short time, is necessary for these plants. For this purpose, an experiment was designed and performed to micropropagate the Singonium ornamental ...
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Introduction: Ornamental foliage plants are commonly used for beautifying indoor spaces. Consequently, determining the best method of mass propagation in a short time, is necessary for these plants. For this purpose, an experiment was designed and performed to micropropagate the Singonium ornamental plant.Materials and Methods: For the first stage, the effect of different plant growth plant growth regulators on the regeneration of nodule explants was evaluated. In this experiment, 1 cm nodal explants were placed in semi-solid MS culture medium containing different concentrations of BA and Kin (0, 1, 2, 3 and 4 mg/l) in combination with 0.2 mg/l IAA. This experiment was performed as a factorial based on a completely randomized design with four replications. The first factor included different types of cytokinin (BA and Kin) and the second factor included different concentrations (0, 1, 2, 3 and 4 mg/l). In the second stage, the effect of different types of culture medium and its different concentrations on plantlet proliferation was investigated. This study was performed as a factorial experiment based on a completely randomized design with five replications. The first factor included the type of culture medium (MS and B5) and the second factor was the different concentrations of culture medium (0.5 and 1). In the third stage of the experiment, the acclimation of in vitro plantlet was investigated. This experiment was performed as a completely randomized design with three replications. The experimental treatments included different culture media: vermiculite, peat moss, vermiculite + peat moss, perlite + peat moss, perlite, coco peat + vermiculite, coco peat + perlite, coco peat and rock wool.Results and Discussion: According to the results of the first experiment, it was found that the use of cytokinin in MS culture medium is effective on plant regeneration. The highest number of regenerated plantlet was observed in culture medium containing 4 mg/l BA with 0.2 mg/l IAA. It is noteworthy that with increasing BA concentration in the culture medium, the plant height decreased, but in contrast, the number of produced plantlets increased. Benzyl aminopurine (BAP) has been introduced as the most important and effective cytokinin in inducing and increasing branching in plants. Research has shown that the use of external benzyl adenine affects plant growth by affecting plant cells or by controlling the accumulation of a number of cytokinin compounds. As a result, the use of cytokinins in culture medium under in vitro culture conditions is necessary to induce and increase cell division. According to the second experimental results, produced plantlets in MS culture medium had higher height, number of roots and root length compared to B5 culture medium. Results also demonstrated that the use of ½ MS in the propagation stage of this plant is appropriate due to the improvement of growth traits such as plantlet height and root length. The use of optimal and suitable culture medium is very effective in the success of plant micropropagation. In the present study, the use of MS culture medium showed better performance compared to B5 culture medium. The appropriate amount of components, better ion strength and more minerals in this culture medium compared to B5 culture medium are probably the factors influencing its superiority in in vitro culture. In the acclimation stage, the use of vermiculite + peat moss culture medium led to 100% adaptation of plants and improvement of growth traits in the plant.Conclusion: The results showed that the type and concentration of cytokinin had a significant effect on the most of the evaluated parameters. With increasing the concentration of cytokinin in the culture medium, the number of regenerated plantlet was increased, but on the other hand, the height of regenerated plantlet also decreased. The use of culture medium containing BA compared to KIN had a more pronounced effect on increasing the number of regenerated plantlets. Application of MS medium compared to B5 medium was more effective in increasing plantlet height, root length, number of roots and number of produced plantlets. Also, half-strength MS medium, increased plantlet height and root length. Therefore, at this stage of propagation, the application of ½ MS culture medium is recommended. In the acclimation stage, it can be stated that the use of vermiculite substrate in combination with peat moss is a suitable option considering 100% compatibility of syngonium plantlets and improvement of growth traits in comparison with other substrates.
Akram Amiri; Mahmood Shoor; Mina Taghizadeh; Seyyed Hossein Nemati; Ali Tehranifar
Abstract
Introduction: The genus of African violet is the most common genus known among the plants of Gesneriaceae family. This genus has beautiful leaves and flowers that are zygomorph. African violet (Saintpaulia ionantha) is a famous ornamental plant due to its various colors and shapes that an excellent model ...
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Introduction: The genus of African violet is the most common genus known among the plants of Gesneriaceae family. This genus has beautiful leaves and flowers that are zygomorph. African violet (Saintpaulia ionantha) is a famous ornamental plant due to its various colors and shapes that an excellent model system for in vitro regeneration studies because of its tissue culture amenability.Besides the importance of this plant in plant production industry and increasing need for new plant varieties with desired traits, more studies for creating African violet with new features by increasing the ploidy level under the in vitro conditions were not done until now. Polyploidy is a widespread phenomenon in the evolution of flowering plant and a key in plant speciation and diversification. Polyploid plants have been used in plant breeding programs for developing superior varieties and restoring the fertility of interspecific or intergeneric hybrids. Polyploidy in ornamental crops were successfully obtained under in vitro conditions. The chemical colchicine can be used as the most effective substance to obtain polyploid plants. In vitropolyploidization has number of advantages such as treatment of more plants with less material, control of test conditions, lower toxicity of chemicals and high success rate. This study with the aim of polyploid induction was carried out by different concentrations of colchicine at various periods under in vitro conditions.Materials and Methods: The best treatment for the shoot regeneration and proliferation was MS medium with 2 mg/l BA after optimizing the tissue culture process. The study performed as follows: plantlets grown under in vitro in proliferation stage were treated with colchicine. The experiment was carried out in a 2-factorial manner based on a completely randomized design and factors were colchicine concentration at 0.02, 0.05, 0.1% and treatment duration for 18, 24, 48 hours. Evaluated characters of regenerated plants were as follows: plantlets survival percentage (the first month, the second month and the third month), morphological traits (include leaf number, petiole length, leaf length, leaf width, petiole diameter, leaf thickness and variation in form, vein, tip and margins leaf), reproductive traits (flower diameter (mm), number of petal, number of flower in inflorescence, height of inflorescence (mm)), microscopic epidermal cells in samples of diploid and tetraploid and ploidy levels. Assessment of flow cytometery was also used for all of the treated plants with colchicine and some diploid control plants and were expressed in the form of the percentage of diploid, mixoploid and tetraploid plants of African violet.Results and Discussion: Results indicated that tetraploidy induction successfully was changed different morphological and cytological characteristics. Plantlets of treated with 0.02% colchicine in all three times only survived after three months, and with increasing treatment time, plantlets survival percentage was reduced. Thus 0.02% colchicine treatment for 24 hour found effective in inducing in vitro culture polyploidy of African violet. Comparison of the leaves of tetraploid with diploid plants indicated that the leaves of tetraploid plants in length and width were bigger than the diploid ones. The selected tetraploid plants showed a more compact shape than the control plants. Other results revealed that the treated plants with colchicine showed delayed growth. The assessment of microscopic indicated that the epidermal cells of tetraploid were larger than diploid. The results of flow cytometery evaluation showed that with increasing duration of colchicine, the number of diploid plants reduced and were added to mixoploid and tetraploid plants. Most plants were mixoploid.Conclusion: The results of this study showed that the diploid and tetraploid plants indicated significant differences in term of morphological traits like leaf number, petiole length, leaf length, leaf width, petiole diameter, leaf thickness and variation in form, vein, tip and margins leaf, reproductive traits like flower diameter, number of petal, number of flower in inflorescence and also size of epidermal cells and flow cytometery evaluation. These differences can be suitable criterion for separating diploid and polyploid plants from each other. Generally, the combination of tissue culture methods and tetraploidy induction can be used as a rapid strategy for achieving new forms and properties of regenerates in vitro on African violet.
Ahmad Sharifi; Seyyedeh Mahdiyeh Kharrazi; Fatemeh Keykha Akhar; Abdolreza Bagheri; Elahe Samari; Maryam Moradiyan
Abstract
Introduction: Gerbera is one of the most important ornamental plants in the world. The importance of Gerbera is due to its beauty, diversity and economically aspects. Traditional propagation methods such as crown division and cutting methods are not suitable for obtaining disease free plants and rapid ...
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Introduction: Gerbera is one of the most important ornamental plants in the world. The importance of Gerbera is due to its beauty, diversity and economically aspects. Traditional propagation methods such as crown division and cutting methods are not suitable for obtaining disease free plants and rapid multiplication. These methods also do not have the capacity to fulfill global demands. Therefore, obtaining efficient protocol for micropropagation of this ornamental plant is necessary.
Materials and Methods: In this study the effect of various factors on in vitro regeneration, proliferation, rooting and acclimation of gerbera capitulum explants were analyzed in four separate experiments. Capitulum explants were first washed with running tap water for 30 min then surface sterilized by dipping in 1.5% sodium hypochlorite solution for 15 min and rinsed with sterile distilled water, followed by immersing in 0.1 % mercuric chloride solution for 10 min. To remove mercuric chloride residue, capitulum was rinsed with sterile distilled water. Subsequent washing was done with sterile distilled water for three times. Sterilization steps were done under laminar air flow hood. For regeneration, eight genotypes of gerbera capitulum explants (Famous, Sunway, Red Pearl, Pink Snow, Popov, Balance, Dune, Eagle)were cultured on solid MS medium containing several cytokynins, BA, TDZ, 2IP or KIN (4 mg/l) in combination with IAA (0.2 mg/l). In proliferation stage, the effect of different concentrations of BA was evaluated on proliferation rate of Sunway regenerated explants. In the rooting stage, Sunway genotype plantlets were cultured on ½ MS medium containing NAA, IBA or IAA (1 mg/l) or ½ MS medium without any hormones. The pH of the medium was adjusted to 5.7-5.8 prior to autoclaving (15 min at 121 oC and 1.5 kg.cm-2 pressure). The cultures were incubated in a growth chamber at 25±2 oC with a 16-h photoperiod (2500-3000 Lux) provided by cool-white fluorescent lamps. For acclimation of rooted plantlets, different substrates used as follow: 1- perlite, 2- perlite: Cocopeat, 3- Cocopeat: peat moss, 4- Cocopeat: peat moss; treated with fungicide.
After 30 days, the response of explants was evaluated for each experiment. Data preparation was done in the Excel program and data analysis was done using JMP-8 software. Mean comparison of the treatments was done by Tukey test and finally the charts were drawn using the Excel program.
Results and Discussion:The results of regeneration stage showed that application of MS media containing kinetin or 2IP did not make an appropriate response to capitulum explants and no regeneration was observed in this condition. The medium containing 4 mg/l BA and 0.2 mg/l IAA indicated the highest percentage of regeneration in all genotypes.
The highest regeneration was observed in Sunway genotype with an average of 21.96%. On the other hand no regeneration was observed in Eagle genotype. In terms of the number of regenerated plantlet, the highest number (61.2) was attributed to the Sunway genotype while no plantlet was recorded for Eagle genotype. No significant differences were also observed between Pink Snow and Dune genotypes.
For the proliferation stage, only Sunway genotype was utilized due to its vigorous growth in comparison to other genotypes. In this stage, the highest (6 regenerated plantlets) and the lowest (1 regenerated plantlet) regeneration rate were observed in MS medium containing 2 mg/l BA and hormone-free medium, respectively. Hormone-free ½ MS medium and ½ MS medium containing 1 mg/l IAA or IBA, indicated the highest rooting rate (100% rooting) while medium containing 1 mg/l NAA showed 55% rooting rate. It seems that the application of NAA in the medium composition had the lowestimpact on the rooting of regenerated plantlets. At the end of the experiment, the highest (90.42%) and the lowest (47.5%) acclimation rate was obtained in peat moss + cocopeat + fungicide medium and perlite medium, respectively.
Conclusions: Generally, for shoot induction of gerbera through capitulum culture, application of MS medium containing 4 mg/l BA and 0.2 mg/l IAA is recommended. It is also concluded that for proliferation stage, the MS medium containing 2 mg/l BA showed the highest rate of regeneration. Using of Hormone-free ½ MS medium is economically affordable. Finally for acclimation of the plantlets, application of peat moss + cocopeat + fungicide medium is recommended.
Ahmad Noroozi; Abdolreza Bagheri; Nasrin Moshtaghi; Ahmad Sharifi
Abstract
Introduction: Anthurium is a popular genus of the Araceae (order Spathiflorae).The flower consists of a protruding spadix containing numerous florets, subtended by a brightly colored modified leaf, the spathe. Anthuriums are bisexual and protogynous.Anthuriumscherzerianum as the most important species ...
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Introduction: Anthurium is a popular genus of the Araceae (order Spathiflorae).The flower consists of a protruding spadix containing numerous florets, subtended by a brightly colored modified leaf, the spathe. Anthuriums are bisexual and protogynous.Anthuriumscherzerianum as the most important species ofAnthurium genus is a potted perennial plant. Due to having beautiful, attractive and long-life flowers, A. scherzerianum can be used for the production of pot and cut flowers. Tissue culture is suggested as the most commonly method in order to rapid propagation and removing disease in a short period of time. This method also recommended for Anthuriumbecause of problems in classical propagation method of this flower..The three basic propagation methods for Anthuriumare propagation by seed, traditional vegetative and tissue culture.Micropropagation of Anthurium is using forcommercial production.
Materials and Methods: In this study, the effect of plant growth regulators and explants on indirect regeneration of A. scherzerianumdetermined in separate experiments. In the first experiment, callogenesis was done by leaf explants on MS medium containing growth regulators, BA in three concentrations (0.5, 1.25 and 2 mg/l) in combination with
2, 4-D (0.5, 1.25 and 2 mg/l) or NAA (0.5, 1.25 and 2 mg/l) and the combinations of TDZ (0.5, 1.25 and 2 mg/l) with
2, 4-D (0, 0.5 mg/l). In the second experiment, regeneration was done on MS medium containing 0.75 mg/l BA with 0.05 mg/l 2, 4-D and 0.1 mg/l NAA and also in combination with TDZ (0.75mg/l). For rooting, MS medium containing different concentrations of IBA and IAA (0, 0.2 and 1 mg/l) were used. Callus induction, regeneration and rooting experiments were done based on completely randomized design, with 12, 6 and 6 replications, respectively.Data from all the schemes used in this study were analyzed with SAS statistical software. The comparison of means using Duncan's multiple range test was evaluated at the 5% level.
Results and Discussion: Analysis of variance showed that the effect of explant type and hormone combinations was significant on the percentage of callogenesis, callus volume and survival percentage. The interaction effect of explant type and combination of hormones was also significant on percentage of callogenesisand the volume of callus. Means comparisons showed that the highest callogenesis, viability and callus volume were achieved on MS medium containing 2 mg/l of BA and 0.5 mg/l of 2, 4-D. Petiole explants, also produced the highest percentage of callus (95%), survival rate (96%) and callus with dimensions of 6 mm2. Callus formation in leaf vein explants was higher than others. The effect of explant type and hormone combinations on regeneration, number of branches, number of leaves and leaf length was significant.The interaction of explant and hormone combinations on regeneration, number of branches, number of leaves and leaf length was also significant. Moreover, results of regeneration experiment indicated that the maximum number of shoots (6.9) and the maximum shoot length (5 cm), number of leaves (18) and the leaf length (2.8 mm) were achieved in 0.75 mg/l BA mg/l of and 0.05 mg/l 2, 4-D. In this study, petiole explants were also regenerated earlier than leaf explants.The effect of hormone combinations and concentrations was significant on rooting specially on the number of roots and root length.Furthermore, results of rooting experiment revealed that the highest rooting percentage (95%), the maximum number of roots (4.5 per plantlet) and the longest roots (3.5 cm) were produced in the medium containing 0.2 mg/l of IBA. Finally, the rooted plantlets were adapted (90%) in vivo condition by placing them on a mixture of cocopeat and perlite (2:1) substrate.
Conclusion: In this study callugensis, regeneration and rooting of A.scherzerianum’s petiole and leaf explants were studied and different levels of plant growth regulators used for callugensis and regeneration. In this study petiole explants showed the highest callugenesis and regeneration. MS medium containing BA (2 mg/l) and 2, 4-D (0.5 mg/l), was the best for callugenesis. Also the highest percentage of regeneration was observed in medium containing BA (0.75 mg/l) and 2, 4-D (0.05 mg/l). Moreover low concentration (0.2 mg/l) of auxin has a better effect on rooting than high levels (1mg/l) so that the highest rooting percentage was produced in medium containing IBA (0.2 mg/l) and the lowest rooting percentage was produced in medium containing IAA (1 mg/l). Anthurium plantlets acclimized is cocopeat and perlite substrate (2: 1) with 90% acclimation.
A. Khazaei; N. Moshtaghi; Saeid Malekzadeh Shafaroudi; Kamal Ghous
Abstract
Introduction: Jujube (Ziziphus jujuba) is one of the most important fruit trees in Asia which has been planted from 3,000 years ago in China for medicinal purposes. Jujube belongs to the Rhamnaceae family. The Jujube fruit is used in fresh and dry forms. The fruit is full of vitamin C and has anticancer ...
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Introduction: Jujube (Ziziphus jujuba) is one of the most important fruit trees in Asia which has been planted from 3,000 years ago in China for medicinal purposes. Jujube belongs to the Rhamnaceae family. The Jujube fruit is used in fresh and dry forms. The fruit is full of vitamin C and has anticancer and medicinal effects. This tree can grow on salty and dry lands in Iran. Therefore, increasing the cultivation area of Jujube can be effective for soil conservation. In the last 20years, cultivation of Jujube is is considerable in Iran specially in the South Khorasan Province and 98 % of total production of Jujube in Iran belongs to this province. The low rate of seed germination and low production of shootlets are the most important problems in Jujube proliferation, so micropropagation of this plant through tissue culture was considered.
Materials and methods: In this study, Cangan ecotype of Jujube was used for multiple shoot regeneration. At the end of May, apical buds of shoots were cut from mature trees of the Research Collection of Jujube at Sarbishe, Birjand, South Khorasan Province in Iran. The buds were disinfected with 70% ethanol for 1 min and 2% sodium hypoclorite for 25 min. Then the buds were rinsed with distilled water for 25 min completely. Apical buds were placed on Murashige and Skoog (MS) medium supplemented with different concentrations of BA (0.5, 1, 1.5, 2 mg/L) in combination with IBA or NAA (0, 0.1, 0.2, 0.4 mg/L). After one month, the shoots with 3-5 cm length were transferred to rooting media (1/2 MS + IBA or IAA : 0.5, 2, 5, 10 mg/L). The data were recorded after shooting and rooting and were analysed in the facorial experiment.
Results and Discussion: The results of variance analysis and mean comparisons showed that there are differences between different levels of IBA and BA alone for the number of shoots and their length (P
