Breeding and Biotechnology of Plant and Flower
Fatemeh Keykha Akhar; Abdolreza Bagheri; Nasrin Moshtaghi; Masud Fakhrfeshani
Abstract
Introduction
Flower color is one of the most significant characteristics in ornamental plant breeding. New varieties of various plants in relation to their flower color have been obtained by monitoring the expression levels of genes involved or regulating the flavonoid and anthocyanin biosynthesis ...
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Introduction
Flower color is one of the most significant characteristics in ornamental plant breeding. New varieties of various plants in relation to their flower color have been obtained by monitoring the expression levels of genes involved or regulating the flavonoid and anthocyanin biosynthesis pathway. Flavonoids possess significant and diverse biological functions. They are the major pigments for flowers, fruits, seeds, and leaves. They are natural products that contain a C6-C3-C6 carbon framework and are synthesized by a branched pathway that yields both colored and colorless compounds. The gene encoding chalcone isomerase (CHI) is among the genes and enzymes identified in the flavonoid pathway. This enzyme catalyzes the isomerization of naringenin chalcone into the corresponding flavanone. CHI enzyme belongs to the family of isomerases, specifically the class of intramolecular lyases. Chalcone isomerase has a core 2-layer alpha/beta structure and has attracted much attention recently due to its role in stress response and pigment production. One of the most effective methods of genetic engineering is the reduction of flower pigments by suppression of required enzymes for their biosynthesis. RNA interference (RNAi) has provided the tool for the investigation of genes involved in the production of flower color. Silencing of any gene in the anthocyanin biosynthetic pathway can result in reduced or inhibited anthocyanin production. RNAi technology is an effective gene silencing method and a powerful tool for studying gene function and development of new traits by transformation of viral RNA or hairpin RNA (hpRNA) constructs into plants. The processing of dsRNA into 21-23-nt small interfering RNAs (siRNAs), and the mediators of RNAi, triggers cognate mRNA degradation. The hpRNAi methodology simply requires a transgene construct containing an inversely-repeated sequence of the target gene flanked with a promoter and terminator which effectively function in plants.
Material and Methods
In this research, with the design and construction of chiRNAi, the transformation of the RNAi construct was carried out of Petunia plants. Potted plants of P. hybrida were grown under standard greenhouse conditions (16-17°C night temperature and 21-24°C day temperature and photoperiod 16/8 (light/dark)). The RNAi construct including the 530 bp cds of the chalcone isomerase (chi) gene and 741 bp of pdk gene as intron between chi sense and antisense were used for transient RNAi-induced silencing. The pBI121-chi530 plasmids were introduced into A. tumefaciens strain LBA4404 by electroporation method. Colonies of A. tumefaciens carrying the desired plasmid were screened by PCR with specific primers for chi gene. RNAi construct co-cultured with petunia’s leave. Samples was kept in dark condition for 3 days and then transferred to branch induction media. Samples were investigated for phenotypical changes and chi gene expression by qRT-PCR.
Results and Discussion
Transgenic lines showed a reduced number of pigments and a faded flower color. So that, in purple petunia, was shown 5 phenotypical groups. These groups was indicated different levels of chi gene silencing. In pink petunia was seen two groups of phenotypical changes. In these plants, chi-RNAi construct was reduced pigment production and so, these plants had faded colors in petals. Also, the chi gene expression was reduced in all transgenic lines. Generally, the results of this research showed that RNAi can be used as an efficient method for gene silencing. The application of gene silencing can indicate the gene’s function in biosynthesis pathways of various components such as anthocyanins. In addition, the chalcone isomerase gene was identified as one of the effective genes in anthocyanin biosynthesis pathway in Petunia plants that could be involved in the production of color in these plants; hence, chi gene silencing resulted in clear phenotypic alterations in this plant.
Conclusion
In general the concentration of the target mRNA in a particular tissue could be a factor that influences silencing efficiency. At very low levels of gene expression, small amounts of the silencing target, mRNA, could be completely degraded by the RNA-induced silencing complex (RISC), whereas the presence of higher amounts of the target mRNA may result in incomplete silencing, allowing some residual functional mRNA to be translated into the corresponding protein. This research demonstrated the hpRNA construct has been successfully established for floral tissues of P. hybrida. The hpRNA construct was developed for chi-RNAi silencing of one of the key genes in the anthocyanin biosynthetic pathway in Petunia flowers. The silencing of the chi gene is a prototype for the modification of the anthocyanin biosynthetic pathway in Petunia through gene suppression. This strategy could also be useful for rapid functional analysis of other genes involved in flower development.
Hoda Zare Mirakabad; Mohammad Farsi; Saeid Malekzadeh Shafaroudi; Mehrdad Iranshahi; Abdolreza Bagheri; Nasrin Moshtaghi
Abstract
Introduction: There is a growing body of the literature that recognizes the importance of ferutinin (C22H30O4) as one of the natural phytoestrogens with potency to treat osteoporosis and some kind of cancers. One of the greatest challenges is availability of ferutinin that is found just in root of some ...
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Introduction: There is a growing body of the literature that recognizes the importance of ferutinin (C22H30O4) as one of the natural phytoestrogens with potency to treat osteoporosis and some kind of cancers. One of the greatest challenges is availability of ferutinin that is found just in root of some plants of the genus Ferula (Apiaceae), which is the reason of high price of it in global markets. Ferula ovina , an endemic plant of Iran, is known as one of the greatest sources of ferutinin. Unfortunately, access to ferutinin requires uprooting Ferula ovina especially older plants with more secondary metabolites content. There is a large volume of published studies describing the importance of tissue culture in propagation of endangered plants including secondary metabolites without possibility of chemical production and with deep dormancy seed exactly like the characters of F. ovina. Up to now, far too little attention has been paid to importance of tissue culture in accessing ferutinin without degrading the germplasm resources of it. The main aim of this study was to find an approach to access ferutinin associated F. ovina germplasm conservation.
Materials and Methods: In this experiment, the aerial parts of F. ovina were collected from Zoshk area (Mashhad, Iran). The plants were recognized as F. ovina by the Institute of Plant Sciences (Ferdowsi University of Mashhad). Tissue culture part was performed with preparation of sterilized media and explants. Therefore, the MS salts and vitamins was applied as basic medium, however MS salt was decreased to half strength for rooting of shoots. The node (root junction) explants were cultured on 24 callus induction/shooting media with different combinations of plant growth regulators (BAP, IAA, Kin, NAA and IBA). The shoots from direct and indirect regeneration were then transferred to media with different combinations of PGRs (BAP, IBA, IAA and NAA) in order to find rooting medium. Following these treatments, unbalanced ANOVA for analysis of data were performed using IBM SPSS 19. The final stage of the study comprised a TLC test for the purpose of finding ferutinin in samples which resulted from tissue culture. For this purpose, air-dried parts of samples were powdered and extraction was done after being in 3-5 times dichloromethane for 24hours. Then after optimizing solvent system was done with selection the ratio that presented bands in middle of TLC paper.
Results and Discussions: The results indicated that from 24 tested media, just 8 of them had potency of callus formation, but just L6 on MS medium containing 1 mg/l BAP and 1.5 mg/l IAA showed significant difference for percentage of callus induction at 5% level with compact green and the heaviest calluses. Although direct and indirect shoot regeneration was observed in this study, L9 on MS medium along with 1.5 mg/l IAA and 3 mg/l kin demonstrated significant difference for percentage of shooting at 5% level. Moreover, R6 on ½MS with 1 mg/l IBA showed significant difference for percentage of rooting of shoots at 5% level. The most surprising observation to emerge from the data comparison was L24 on half strength MS medium containing 0.2 mg/l BAP and 3 mg/l IBA with potency of callus induction, shooting and rooting of shoots. Regarding to the results of TLC test, ferutinin positive bands were observed in some samples.
Conclusions: The aim of the present research was to examine possibility of achieving ferutinin without need to uproot F. ovina because there are several problems to achieve this valuable sesquiterpene: 1) Chemical production of ferutinin is impossible, 2) It could be accessible just from roots of genus Ferula, 3) Propagation with seeds of F. ovina is limited because of morphophysiological dormancy of them, 4) Natural habitat of this plant in Iran is going to be destructed, 5) Access to natural habitat is difficult, 6) Time of access is limited with short growth season, 7) Maintaining F. ovina in greenhouse condition is impossible. The results of this research support the idea that producing ferutinin in Iran without any harmful effect on germplasm resources of F. ovina is possible. This is the first study of high scale commercial production of ferutinin which examine associations between tissue culture and ferutinin production.
Maria Beihaghi; Abdolreza Bagheri; Seyyed Hassan Marashi; Mojtaba Sankian; Afsaneh sadat Farsad
Abstract
Introduction: Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition and widely used to produce clones of a plant in a method known as micropropagation. Plant research often ...
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Introduction: Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition and widely used to produce clones of a plant in a method known as micropropagation. Plant research often involves growing new plants in a controlled environment. These may be plants that we have genetically altered in some way or may be plants of which we need many copies all exactly alike. These things can be accomplished through tissue culture of small tissue pieces from the plant of interest. These small pieces may come from a single mother plant or they may be the result of genetic transformation of single plant cells which are then encouraged to grow and to ultimately develop into a whole plant. Tissue culture techniques are often used for commercial production of plants as well as for plant research. Tobacco (Nicotiana tabacum L.) is one of the most important model plants used in the physiologic, genetic and tissue culture studies. The manipulation of tobacco genetic structure requires an efficient technique of gene transferring and regeneration. Whereas, the tobacco plant is a very effective bioreactor in the production of recombinant proteins, in this research we optimized the best tissue culture system and also, genetic transformation process of this plant.
Materials and Methods: Our plant tissue culture protocols, Include helpful information for Murashige and Skoog media, plant growth regulators, plant growth hormones, plant transformation systems, and other products for plant tissue culture. For this purpose, different concentrations of sucrose and 4 combinations of growth regulators (BAP and NAA) on callus induction, direct shoot regeneration and rooting were examined in a factorial experiment based on completely randomized design with 3 replications. The sensitivity of tobacco explants to kanamycin was examined through the cultivation of them on the selective medium with different concentrations of antibiotic. For genetic transformation, agrobacterium tumifacious (GV3101) harboring plasmid pBI121 was used and the transgenic plants were confirmed by PCR analysis.
Results and Discussion: The results of variance analysis and the means comparison showed that the best medium for callus induction was M1 (0.1 mg/l NAA and 1 mg/l BAP) with 15 g/l sucrose in the leaf explants, while the most direct shoot regeneration rate was obtained on the M1 medium with 30 g/l sucrose concentration. High-frequency of rooting was also influenced by 0/1 mg/l NAA and 60 g/l sucrose. So, supplementing the medium with NAA and BAP at different concentrations facilitated induction of multiple shoots from explants. NAA was proved to be the best and the number of shoots increased with increase in the concentration up to (0.1 mg/l), and exceeding this concentration resulted in decline in percent response as well as number of shoots was recorded shoot regeneration. The concentration of BAP was further increased a linear increase in the number of shoots was observed up to an optimal level (1 mg/l). Beyond the optimal concentration (1 mg/l), a decrease in the response as well as number of shoots was recorded due to profuse basal callusing. The effect of cytokinins on multiple shoot regeneration, higher concentrations of NAA found to be inhibitory for shoot regeneration because of huge callusing which hampered the growth and development of new shoots. Also different concentrations of sucrose have a different effect on the shoots and callus. The concentration of sucrose had significant effect on direct shoot regeneration. The main effect of sucrose concentration, concentration of 30 grams per liter, compared with a concentration of 15 grams per liter had the highest direct shoot regeneration. Concentration of 50 mg/l kanamycin could completely prevent the regeneration of untransformed explants so was used in the selective culture medium. Subsequently, the presence of nptII gene (798 bp) in the transgenic plants was confirmed and the transformation efficiency obtained by using the agrobacterium-mediated transformation was more than 95%.
Conclusions In present research, an efficient in vitro regeneration protocol has been developed for tobacco, where different factors including the age of the explant and plant growth regulators were optimized for maximum propagation of tobacco. The results showed that regeneration and transformation method described here is highly efficient and fast for the introduction of any foreign gene directly in tobacco plant.
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 ...
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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.
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.
Seyyedeh Mahdiyeh Kharrazi; Ali Tehranifar; Seyyed Hossein Nemati; Abdolreza Bagheri
Abstract
Introduction: Amaryllis is grown as pot outdoor plant and cut flower. Generally, this ornamental plant propagates by seed, suckers and scale cutting. Propagating by seed is not commercial and often used to produce new varieties. On the other hand, number of bulblets per mother bulb is very low under ...
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Introduction: Amaryllis is grown as pot outdoor plant and cut flower. Generally, this ornamental plant propagates by seed, suckers and scale cutting. Propagating by seed is not commercial and often used to produce new varieties. On the other hand, number of bulblets per mother bulb is very low under normal condition. Besides each bulb produces only 2 or 3 bulblets in a growing season and they become mature and produce flower stalk after 2 to 3 years. In some cases bulbs have no capacity to produce bulblet. Therefore, one of the strategies for shortening the growth period of the plant is to improve the traditional methods of plant propagations.
Materials and Methods: This study was done as a factorial experiment in a completely randomized design with 7 replications to evaluate the effects of medium and position of twin scales in mother bulbs on propagation of bulblets, in order to increase the rate of propagation of this ornamental plant. To measure wet and dry weight of explants, 3 replicates were used. For propagation, bulbs were cut radially into 12 equal pieces, so that each pieces were contained a part of the basal plate. To evaluate the effects of position of twin scales in mother bulbs, pieces were divided as twin scales and classified in 5 groups, so that the outermost twin scales was grouped in class 1 and the innermost twin scales was grouped in class 5. After that, the scale cuttings were dipped in 0.1 % carbendazim solution for 25 minutes and then surface water were dried using sterilized tissue paper. Media that used in this study were sand, perlite, vermiculite, Peat moss and cocopeat. For removing possible contamination from the media, all media were autoclaved for 30 minutes at 121 °C. Then twin scales cuttings were cultured in vented transparent plastic containers that filled with different media and were kept in a growth chamber at 25 °C and 16 hours lighting.Number of produced bulblet, bulblet diameter, root number, root length, fresh and dry weight of plants and browning rate of scales were recorded at the end of the experiment.
Results and Discussion: The results showed that medium and twin scale position in the mother bulb had a significant effect on the quality produced bulblet. The highest fresh weight of bulblet (1.58 g), bulblet dry weight (0.21 g) and the maximum diameter of the produced bulblet (1.5 cm) were obtained in the outermost twin scales and peat moss medium. Analysis of variance showed that the effect of culture medium on the number and length of produced leaf was significant (p
Seyyedeh Mahdiyeh Kharrazi; Seyyed Hossein Nemati; Ali Tehranifar; Abdolreza Bagheri; Ahmad Sharifi
Abstract
Carnation is considered as the world's third most important cut flower. Tissue culture techniques offer suitable method for micropropagation of this ornamental plant. However, one of the problems during in vitro culture of carnation is vitrification. Ratio of ammonium to nitrate and agar concentrations ...
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Carnation is considered as the world's third most important cut flower. Tissue culture techniques offer suitable method for micropropagation of this ornamental plant. However, one of the problems during in vitro culture of carnation is vitrification. Ratio of ammonium to nitrate and agar concentrations in the medium affect this phenomenon. Therefore, in this study the effect of these factors on the rate of proliferation and the vitrification of carnation (Dianthus caryophyllus L.) cultivar Innove Orange Bogr, was evaluated. In this investigation lateral buds were cultured on MS medium containing 1 mg/l BA, 0.1 mg/l NAA and with different concentrations of agar and different ratio of ammonium to nitrate. The results showed that increasing in the agar concentration to 12 g/l lead to decreasing the rate of vitrification but regeneration also declined. Increase of agar concentration cause limitation in nutrient absorption by plants. Also, decrease in the ratio of ammonium to nitrate in the medium reduces the amount of vitrification, but did not result in adverse effects on plant regeneration rates. Multiple regressions showed that the effect of ammonium to nitrate ratio on vitrification was higher than agar concentration. So by considering the amount of shoot regeneration and vitrification, to obtain the most normal shoots, the concentration of 10 g/l agar with ammonium to nitrate ratio 1:6, is recommended.
Seyyedeh Mahdiyeh Kharrazi; Seyyed Hossein Nemati; Ali Tehranifar; Ahmad Sharifi; Abdolreza Bagheri
Abstract
Abstract
Carnation (Dianthus caryophyllus L.) is the third most important cut flowers in the world. Tissue culture techniques offer an efficient method for micropropagation of this ornamental plant. In present work effects of Kinetin (Kin) and Benzyl Adenine (BA), on shoot multiplication and hyperhydricity ...
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Abstract
Carnation (Dianthus caryophyllus L.) is the third most important cut flowers in the world. Tissue culture techniques offer an efficient method for micropropagation of this ornamental plant. In present work effects of Kinetin (Kin) and Benzyl Adenine (BA), on shoot multiplication and hyperhydricity of four carnation cultivars (Prado Aquila Kgr, Skimo Mogr, Mondeo Kgr and Innove Orange Bogr) were studied. Explants from nodal segments were cultured on MS medium supplemented with different concentrations of BA (1, 2, 3 and 4 mg/l) and Kin (1, 2, 3 and 4 mg/l) in combination with 0.2 mg/l NAA, 30 g/l sucrose and 8 g/l agar. Rooting of regenerated shoots was done in the MS medium supplemented with 1 mg/l NAA. Results indicated that there is a significant difference among cultivars shoot regeneration, Eskimo and Prado Aquila Kgr with 3.2 and 1.5 shoots show the highest and lowest regeneration rate, respectively. Increasing the concentration of cytokinin from 1 mg/l to 4 mg/l lead to increase regenerated shoot number from 1.7 to 2.4 shoots per explant and increase hyperhydricity from 12% to 54%. In addition high concentration of cytokinin, especially BA, decreased height of regenerated shoots. Based on hyperhydricity percentage of regenerated shoots, there was a significant difference between cultivars and cytokinins. Mondeo Kgr and Prado Aquila Kgr showed highest (44%) and lowest (23%) hyperhydricity, respectively and explants cultured in medium supplemented with BA caused more (40%) hyperhydricity than Kin (26%). Results of present work showed that by increasing the concentration of cytokinin specially BA, multiplication will be increased but also it will increase the hyperhydricity of plantlets and consequently it will lead to death of them. By considering the amount of multiplication and frequency of hyperhydricity for obtaining the highest number of normal shoots, using of MS medium containing 1 mg/l BA in combination with 0.2 mg/l NAA is suggested.
Keywords: Carnation, Benzyl Adenine, Kinetin, Hyperhydricity, Shoot multiplication, Micropropagation
Amirghaffar Ghaffar Shahriari; Abdolreza Bagheri; Ahmad Sharifi; Nasrin Moshtaghi
Abstract
Abstract
Alstroemeria is one of the most important cut flowers in the world which is multiplicated by rhizome explants in in vitro conditions . The major problem in using rhizome explants is fungal and bacterial contamination which inhibit the preparation of strill explants. Therefor in this experiment, ...
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Abstract
Alstroemeria is one of the most important cut flowers in the world which is multiplicated by rhizome explants in in vitro conditions . The major problem in using rhizome explants is fungal and bacterial contamination which inhibit the preparation of strill explants. Therefor in this experiment, the effect of disinfecting compounds like sodium hypochlorite, mercury chloride, nano silver particles, the antibiotics of streptomycin, penicillin, cefotaxime and carbendazim fungicide were examined on controlling of contamination. After disinfection, the explants were cultured on MS medium supplemented with 2 mg/L BA and 0.2 mg/L NAA. Contamination percentage was measured after three weeks. The results showed that the treatment of plants with 0.4% carbendazim fungicide and disinfection by 70% ethanol for one minute and %3 sodium hypochlorite for 20 minutes for caralis cultivar, and disinfection by 70% ethanol for one minute and %3 sodium hypochlorite for 20 minutes follow by culturing in medium with 200 mg/l either of streptomycin and penicillin in Bordeaux cultivar had the lowest contamination percentage. Contamination was not controlled by nano silver particles in studied plants.
Keywords: Alstroemeria, sodium hypoclorite, mercury chloride, antibiotics, carbendazim
Zeynab Ghayoor Karimiani; Abdolreza Bagheri; Maryam Jafarkhani Kermani; Gholamhossein Davarynejad
Abstract
Abstract
Gerbera is one of the most demanded plants in commercial cut flowers plants. Optimizing the micro-propagation of new cultivars is therefore important. In this research “Red Explosion” cultivar was used and Capitola (as explants) were directed to a modified solid Murashigue and Skoog medium ...
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Abstract
Gerbera is one of the most demanded plants in commercial cut flowers plants. Optimizing the micro-propagation of new cultivars is therefore important. In this research “Red Explosion” cultivar was used and Capitola (as explants) were directed to a modified solid Murashigue and Skoog medium with four concentration of TDZ (0.25, 0.50, 0.75 and 1.00 mgL-1) for regeneration. Three concentration of kinetin (4, 6 and 8 mgL-1) were applied and plantlets were transferred to free hormone medium for rooting. Number of new leaves produced, and fresh mass accumulation of cultured plantlets were used to determine the growth rate at 20 and 40 days after culture. Results showed that maximum shoot regeneration was occurred in 0.75 and 1.0 mgL-1TDZ. In proliferation stage first 20 days maximum increase in fresh weight in 6 mgL-1 kinetin and in second 20 days after culture in medium contained 4mgL-1 kinetin were observed. The highest number of new leaves was produced on the medium containing 6 mgL-1 kinetin and 87% of plantlets rooted after one month.
Key words: Gerbera, Tissue culture, Thidiazuron, Kinetin
