Growing vegetables
Fahimeh Yarmohammadi; Alireza Motallebiazar; Samaneh Kazemiani; Mina Amani
Abstract
AbstractIntroduction: Considering the sensitivity of potatoes to viruses, the production of virus-free plants through in vitro cultivation and their propagation leads to a reduction in costs and an increase in yield. One of the effective methods of reducing plant diseases and producing disease-free microtubers ...
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AbstractIntroduction: Considering the sensitivity of potatoes to viruses, the production of virus-free plants through in vitro cultivation and their propagation leads to a reduction in costs and an increase in yield. One of the effective methods of reducing plant diseases and producing disease-free microtubers is the use of in-vitro production methods. Considering the role and importance of macro elements and micro elements in the growth of microtubers, it is possible to change the composition of MS culture medium by changing the concentration of salts of macro elements and micro elements without disturbing the balance of elements. This experiment aims to investigate the effect of different concentrations of macro elements (2 Mac, Mac, ½ Mac) and micro elements (2 Mic, Mic, ½ Mic) of MS culture medium in combination with two concentrations of sucrose (80 and 160 g/liter) was performed on in vitro micronodulation of Agria potato.Materials and Methods: This experiment to investigate the effect of different concentrations of macro elements (2 Mac, Mac, ½ Mac) and micro elements (2Mic, Mic, ½ Mic) of MS culture medium in combination with two concentrations of sucrose (80 and 160 (g/liter) on in vitro microtuberation of Agria potato was carried out as a factorial experiment in the form of a completely randomized design with 3 replications in the plant tissue culture laboratory of the Department of Horticultural Sciences, Faculty of Agriculture, University of Tabriz. Lateral buds obtained from in-vitro shoots were used as explants and were cultured under sterile conditions on different culture mediums for the purpose of microtuberation, and the cultures were kept in continuous darkness and at a temperature of 18±2°C were kept in the growth room. During one month, Microtuber initiation rate and after two months, microtuber formation characteristics were measured.Results and Discussion: The results of the analysis of variance showed that the effect of the concentration of micro elements and the interaction effects of micro elements with different concentrations of sucrose and macro elements were significant only in the case of two traits, the percentage and the speed of microtuber initiation, while all microtuber traits productivity was significantly affected by the interaction of micro elements and macro elements. In all culture mediums with 8% sucrose, the initiation percentage of microtubers was 100% and the initiation rate was also maximum. However, the highest percentage of microtuber formation, weight, length, diameter and number of buds on microtuber was obtained in 2Mac culture medium with 16% sucrose. The results showed that the microtuber that had more weight and size had a higher percentage of dormancy and the buds on the microtuber were not able to germinate and produce microtuber during the stages of microtuber formation.Conclusions: In all traits related to microtubers, except for percentage and speed of microtubers initiation, the effects of micro elements, macro elements and sucrose elements were not significant, and this shows that the three investigated factors cannot independently improve microtubers formation is effective in Agria variety. In all traits of micronodulation, the interaction effect of low consumption elements with other two factors was not significant and this shows that the concentration of low consumption elements in Agria variety is not critical for micronodulation. In all culture mediums with 8 % sucrose, the initiation percentage of microtubers was 100 % and the initiation speed was also the maximum, but when double the concentration of macro elements and 16 % sucrose were used, the initiation percentage and the initiation speed of micro-glands in Agria variety showed a significant decrease. The percentage of micro tuber formation, weight, length, diameter and number of buds on the micro tuber in Agria cultivar were significantly affected by the mutual effect of the concentration of macro elements and sucrose, and the 2 Mac culture medium has 16 % sucrose in the first priority and the ½ culture medium Mac with 8 % sucrose in the second priority was better than the other treatments in terms of the investigated traits. In this research, it was found that the produced micro glands with greater weight and size had a higher percentage of dormancy and during the stages of micro glandogenesis, the buds on the microtubers were not able to germinate and produce micro tubers.
Growing vegetables
Farzad Abdollahi; Alireza Motallebi-Azar; Gholamreza Gohari; Bahram Dehdar; Amir Kahnamoii; Fatemeh Shariat
Abstract
IntroductionGrapheneis one of the new carbon nanomaterial that has unique physical properties and potentially important biological applications. Nanosheet Graphene Oxide has shown great potential to improve plant performance in various areas. Microtuber production technology is also used as a tool to ...
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IntroductionGrapheneis one of the new carbon nanomaterial that has unique physical properties and potentially important biological applications. Nanosheet Graphene Oxide has shown great potential to improve plant performance in various areas. Microtuber production technology is also used as a tool to reduce the time needed to produce economic plant resources, increase the quality of seed tubers, and produce microtubers throughout the year. The aim of this study was to evaluate the effect of Nanosheet Graphene Oxide on the improvement of micropropagation and microtuberazation in potato var. Agria under in vitro conditions. Materials and MethodsSingle node explants obtained from in vitro virus-free plantlet (maintained in tissue culture laboratory, Department of Horticultural science, University of Tabriz) were cultured into modified Murashige and Skoog (MS) medium containing four concentrations of Nanosheet Graphene Oxide (0, 25, 50 and 75 mg/L) carried out in the completely randomized design (CRD) with four replications and kept at 25±2 degree centigrade and a photoperiod of 16 hours of light. The proliferation traits such as leaf length, leaf width, plantlet fresh weight, number of leaves and shoots were recorded. Then, single node explants were transferred to Murashige and Skoog (MS) medium with four concentrations of Nanosheet Graphene Oxide (0, 25, 50 and 75 mg/liter) and kept for two months in complete darkness and at 18±2 ºC and microtuber production indices such as microtuber number, diameter, length and weight, microtuberization percentage, shoot length, microtuber with dormancy were measured. Results and DiscussionThe results of analysis of variance showed that different concentrations of Nanosheet Graphene Oxide had a significant effect on all traits in proliferation and microtuberization stages. Among different levels of Nanosheet Graphene Oxide, application of 75 mg/L showed the best response for leaf length, leaf width, and plantlet fresh weight, followed by 50 mg/L for the number of leaves and shoots, and lastly, 25 mg/L for shoot length. At a concentration higher than 50 mg/L (75 mg/L graphene oxide), the number of leaves not only remained constant but also showed a decreasing trend. Effect of different NGO concentrations on the shoot length showed that there was no significant difference between different concentrations of NGO and the shoot length remained constant, but the difference between the control treatment and NGO was significant. The maximum shoot length was obtained at a concentration of 25 mg/l NGO. The different concentrations of NGO had significant effect on all microtuberization traits at 1% probability level. Mean comparison results for different concentrations of NGO showed that the highest value of the microtuber length, diameter and number were obtained at 25 mg/liter NGO. However, all microtuber traits were not increased at above 25 mg/liter NGO. With the increase in NGO concentrations, the yield of microtuber weight and microtuberization rate remain constant, and it is also possible that these traits will decrease significantly with the increase NGO concentration. The highest yield of microtuber weight and microtuberization rate were obtained at the 25 mg/L NOG, and higher concentrations did not increase them. There was a significant difference between different concentrations of NGO and the control treatment in the number of lateral shoots, so that the maximum number of lateral shoots was obtained at a concentration of 25 mg/L of NGO. Also, concentrations above 50 mg/L of NGO had less effect on the number of lateral shoots and with increasing concentration, the number of shoots decreased significantly. The maximum microtuber weight was obtained at high concentrations of NGO. In other words, with the increase of NGO concentration, the microtuber weight increased, and the most effective concentration was 75 mg/L of NGO for this trait. Although all concentrations of NGO are favorable for this purpose, it is possible that the concentration of 25 mg/l is the most NGO concentration. ConclusionThe results of this research showed that the of 50 and 75 mg/L of Nanosheet Graphene Oxide were the best concentrations micropropagation and microtuberization. 25 mg/L of Nanosheet Graphene Oxide was most efficient concentration . Although these experiments were performed without the use of growth regulators, the addition of Nanosheet Graphene Oxide to the medium increased micropropagation and microtuberization. Therefore, Nanosheet Graphene Oxide can be used as a tool for efficient micropropagation and increasing the quantity and quality seed tubers.
Breeding and Biotechnology of Plant and Flower
Mohammad Esmaeil Naddaf; Ebrahim Ganji Moghadam; Gholmreza Rabiei; Abdolrahman Mohammadkhani
Abstract
Introduction Sweet Cherry (Prunus avium) belongs to the Rosaceae family, which due to vegetative propagation problems, in vitro propagation is recommended to increase mass and disease-free production. Micropropagation has many advantages over other vegetative methods. Although the most suitable ...
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Introduction Sweet Cherry (Prunus avium) belongs to the Rosaceae family, which due to vegetative propagation problems, in vitro propagation is recommended to increase mass and disease-free production. Micropropagation has many advantages over other vegetative methods. Although the most suitable organ that preserves the genetic characteristics of the cultivar is bud meristem, plant regeneration from meristem culture is difficult in many species of woody plants, so micro-grafting is a suitable technique to overcome these problems. The aim of this study was to investigate the effect of scion size and origin of commercial sweet cherry cultivars interact with micrografting on the vegetative rootstocks.Materials and Methods In this study, factorial experiment was used as a test unit in a completely randomized design (CRD) with two factors in five replications and ten seedlings per replication. The first factor was cultivar in seven levels (B: Bing, D: Dovomres, H: Haj Yousefi, P: Pishres, S: Siah- Mashhad, T: Takdaneh, Z: Zard) and the second factor was scion type in four levels (M1R1: 2 mm in vivo explant, M2R1: 5 mm in vivo explant , M1R2: 2 mm in vitro explant and M2R2: 5 mm in vitro explant). To prepare the scion, 1.5 to 2 cm long explants were isolated from shoot tips and then disinfected with 75% ethanol and 20% Sodium hypochlorite. After rinsing with distilled water, the shoot tips with 2 and 5 mm length were extracted for in vivo explants. In vitro explants were obtained from shoo tips that was previously established in MS culture medium with supplement of 1 mg.l-1 of BAP. The meristems were prepared in 2 and 5 mm and used as in vitro explant. 5 cm length in vitro shoots of sweet cherry ‘Gisella 6’ was used as rootstock. Micro-grafting was performed according to the standard method for sweet cherries. Micro-grafted plantlets were transferred to MS medium supplemented with 1 mg.l- l BAP, and kept under low light (100 lux) condition for one week, then transferred to growth chamber at 27.1 °C photoperiods 16/8 hrs light/darkness (1500 lux). In order to induce root, grafted plantlets were transferred to Perlite: MS medium supplement with 1 mg.l- l IBA. After rooting, plants were placed in polyethylene pots containing perlite: peatmoss (1:1) for acclimation. Micro-grafting success indices were recorded in each of the micro-grafted plantlets. The data were analyzed by SAS statistical software (9.1) and the means were compared by Duncan's multiple range test (1 and 5 % of probability levels).Results and Discussion The results showed that in all indices there was a significant difference between scion types and cultivar scion type interactions except grafting time, but there was no difference between cultivars (except longitudinal growth of scion). Among the scion types, the 5 mm in vitro scion (M2R2) had the highest micro-grafting success rate (42%), number of leaves (3.7), longitudinal growth (6.3 cm) and taken grafting time (two days). Finally, in successful micro-grafted plants, ‘Pishres’ cultivar had better results in rooting (32.8%) and ‘Zard’ cultivar in acclimation (3.4%) traits. Probably the presence of leaves led to better nutrient supply and surface contact, so it mostly improved the success of micrografting technique. In this study, micro-grafting success indices were lower than previous reports using seedling rootstocks. This might be due to difficult grafting operations, poor rootstock-scion communication, low physiological activity, and high in vitro oxidase activity. In the type of scion, micro-grafting success rate of 5 mm in vitro scions (include leaf primordia), was better than 2 mm scions (without leaf primordia). These results were consistent with most reports in sweet cherries and other stone fruit that were more successful in micro-grafting using larger in vitro explant.Conclusion Based on our results, it can be concluded that the micro-grafting method in sweet cherry micro-propagation is a fast practical method with high potential for production and regeneration of healthy orchards, which is also possible for other cultivars. In micro-grafting success, in vitro explants are preferable to explants taken directly from in vivo mother trees, and the use of larger explants for scion is recommended due to the presence of leaf primordia in micro-grafting success. However, smaller-size explants are more likely to produce healthy plants.
Breeding and Biotechnology of Plant and Flower
Nima Javaheri; Behzad Kaviani
Abstract
Introduction
Lily, a member of the genus Lilium, belonging to the Liliaceae family is one of the most important commercial pot and cut flower species and one of the three major bulb crops in the commercial market because of its large, colorful and fascinating flowers. Lily hybrids are the most ...
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Introduction
Lily, a member of the genus Lilium, belonging to the Liliaceae family is one of the most important commercial pot and cut flower species and one of the three major bulb crops in the commercial market because of its large, colorful and fascinating flowers. Lily hybrids are the most economically important plants with varied flowers. Hybrid Eastern lily (Lilium oriental hybrid ‘Casablanca’) is a perennial bulbous plant that its propagation by bulb in natural condition is time-consuming, so produces 1–2 bulblets per bulb scale in one years’ time which is not sufficient for large scale cultivation of this plant. One of the most important and best methods for vegetative propagation and breeding of lilies is in vitro bulb scale culture. In vitro adventitious bud regeneration from scales of Lilium rely on many factors like cytokinin and auxin concentrations such as BA and NAA. The successful use of tissue culture techniques for rapid propagation of some species of the genus Lilium including L. ledebourii, L. orientalis, L. longiflorum, L. japonicum, L. speciosum, L. concolor, L. nepalense, L. regale, L. oriental hybrid, L. Asiatic hybrid has been reported. The purpose of the current study was to evaluate the effect of different concentrations of BA and NAA on in vitro proliferation of Lilium oriental hybrid ‘Casablanca’ using bulb scale as explant to establish a suitable protocol.
Materials and Methods
Effect of various concentrations of 6-benzyle adenine (BA; 0, 0.5, 1 and 2 mg l–1) and ɑ-naphtaleneacetic acid (NAA; 0, 0.1, 0.2 and 0.4 mg l–1) were evaluated on in vitro proliferation of L. ‘Oriental’. Bulb scale as explant and MS basal medium as culture medium were used. Activated charcoal was applied to inhibit the browning of the culture medium and explant. The experiments were conducted in completely randomized design (CRD). The 16 treatments were applied, each treatment had 4 replications and each replication had 4 individuals. Therefore, in these experiments, a total of 192 bulbs were used. Traits including total plantlets fresh weight, leaf length, leaf number, bulblet weight, bulblet diameter, bulblet number, survival percentage, root length and root number related to in vitro proliferation were measured. All the statistical analyses were done by using SAS and Tukey’s test. Arcsin software was used for changing percent data.
Results and Discussion
The interaction effect of BA and NAA was significant for all measured traits. Results showed that the maximum number of bulblet (8.66) and root (5.36) were obtained in culture medium enriched with 0.5 mg l–1 BA together with 0.4 mg l–1 NAA. Culture media supplemented with 0.5 mg l–1 BA together with 0.2 mg l–1 NAA and 1 mg l–1 BA together with 0.1 mg l–1 NAA with induction of 7.33 bulblets per explant were suitable media. The largest number of leaf (4.33) was measured in culture medium containing 1 mg l–1 BA together with 0.1 mg l–1 NAA. The highest bulblet weight was measured in culture medium supplemented with 1 mg l–1 BA along with 0.2 mg l–1 NAA. The greatest survival rate (100%) was observed in medium enriched with 0.5 mg l–1 BA together with 0.1 mg l–1 NAA. Survival rate (90%) in explants treated with 2 mg l–1 BA along with 0.4 mg l–1 NAA was high. Obtained results revealed that the presence of both BA and NAA in culture media for enhancement of most traits is necessary and critical. Plantlets were transferred to a growing medium containing cocopeat, peat moss and perlite in identical proportion for acclimatization following proliferation. Approximately, 90% of regenerated plantlets survived and were morphologically similar to the mother stocks. This study will help the producers and breeders for commercial and improvement purposes. The effective role of the simultaneous presence of both auxin and cytokinin in the culture medium in effective organogenesis was shown in the present study. Similar findings were reported for some lilies such as L. ledebourii (Baker) Bioss., L. longiflorum and L. regale. Auxin was effective in stimulating bulb production and growth of the aerial part of the eastern lily, and its presence along with cytokinin is essential for leaf induction. Some studies have reported similar results. The type and optimal concentration of plant growth regulators (PGRs) in the culture medium for suitable in vitro propagation varies in different species. Genetic variations (species type), differences in the amount of endogenous production of PGRs and their interaction with each other are among some reasons for this difference. The proper ratio of auxin and cytokinin in the culture medium is effective for inducing cell division, cell differentiation, organogenesis and finally for achieving a complete plant. Root production with appropriate quantity and quality leads to the suitable survival of seedlings resulting from the growth of cultured explants under in vitro conditions and adapted plants. Current study showed that the presence of both BA and NAA is better than the presence of one of these two PGRs for induction and growth of root. Some similar findings were reported, however in most studies, the presence of auxin as individual PGR has been found to be more suitable for root induction.
Seyyedeh Mahdiyeh Kharrazi; Ali Tehranifar; Ahmad Sharifi
Abstract
Introduction: Success in tissue culture technique, especially in bulbous plants, depends on the microbial contamination control during in vitro culture. Applying different treatments, such as heat treatment and usage of fungicides, can control the microbial contamination and consequently increase the ...
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Introduction: Success in tissue culture technique, especially in bulbous plants, depends on the microbial contamination control during in vitro culture. Applying different treatments, such as heat treatment and usage of fungicides, can control the microbial contamination and consequently increase the percentage of explant survival.
Materials and Methods: This study aimed to investigate the effect of heat treatment and fungicide on reducing the contamination during in vitro culture of narcissus. So, an experiment was done as a factorial experiment in a completely randomized design with two factors, including benomyl concentration in the medium (1 and 2 g/l) and heat treatments (two levels, with and without heat treatment), with 10 replications. In order to sterilizing the plant materials, damaged and infected scales were removed firstly and then bulbs were washed for 30 minutes with running tab water and a few drops of dishwashing liquid. For applying heat treatment, bulbs were divided into two groups. In the first group, heat treatment was not applied and in the second group heat treatment 54 °C was applied for one hour using water bath. After this step, bulbs surface were sterilized by dipping in 70% ethanol for one minute and rinsed with sterile distilled water, followed by immersing in 1.5% sodium hypochlorite solution for 30 min. After sterilization with sodium hypochlorite solution, bulbs were washed three times with sterile distilled water under laminar air flow hood. After sterilization step, bulbs were cut into 32 twin scales explants and cultured in MS medium supplemented with 1 mg/l BA and 0.2 mg/l NAA + benomyl (1 or 2 g/l). After 30 days, the response of explants (number of produced bulbs, percentage of explant survival, percentage of bacterial contamination, percentage of fungal contamination, percentage of browning) was evaluated. 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 LSD test and finally the charts were drawn using the Excel program.
Results and Discussion: The results showed that increasing the concentration of benomyl in the medium and applying heat treatment had negative effect on regeneration potential of explants, so that the maximum regeneration mean were observed when heat treatment was not applied for explants and medium contains 1 g/l benomyl. Using the heat treatment and application of 2 g/l benomyl in the medium leads to the lowest regeneration amount. On the other hand, evaluating the browning percentage of explants showed that the effect of treatments was significant in this trait. Applying the heat treatment and using 2 g/l benomyl in the medium had severe effect on the increasing of explant browning and the maximum mean was observed in this treatment. But reducing the benomyl concentration in the medium and none application of heat treatment caused the lowest amount of explant browning. Contamination percentage that includes bacterial and fungal contamination is an important parameter in this study. Explants that cultured in the medium containing 1 g/l benomyl and applying heat treatment showed the highest contamination percentage, which contains 21% fungal and 14% bacterial contamination. The lowest percentage of contamination was observed when heat treatment applied and medium contains 2 g/l benomyl. However, this treatment caused the highest percentage of explant browning that lead to reduction of explants regeneration potential. Researches showed that the use of fungicides can help to control tissue culture contamination and according to previous studies, benomyl is the most effective treatment against fungal infection. As benomyl is considered as a systemic fungicide, so it is useful to eliminate the internal fungus. On the other hand, there are some reports about the positive effect of heat treatment on the control of tissue culture contamination. As regards this investigations were done in 1914 to 1997 and then stopped, so it seems that application of this treatment had no sufficient efficiency for contamination control during in vitro condition.
Conclusion: Therefore, by considering the browning, regeneration and contamination percentage, non-application of heat treatment and usage of 1g/l benomyl fungicide in the medium for in vitro culture of Narcissus twin scales explants is recommended.
Mahnaz Sayadi Nejad; Seyyed Mostafa Sadeghi
Abstract
Introduction: Zamiifolia is a perennial ornamental plant and is one of the most important medicinal plants of the Aracea family. The origin of this evergreen, low-anticipated plant is East Africa. Zamiifolia spreads through the leaves and split rhizomes, which is very time-consuming. The traditional ...
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Introduction: Zamiifolia is a perennial ornamental plant and is one of the most important medicinal plants of the Aracea family. The origin of this evergreen, low-anticipated plant is East Africa. Zamiifolia spreads through the leaves and split rhizomes, which is very time-consuming. The traditional Zamiifolia proliferation method have been done by dividing rhizomes and leaf cuttings, but the production efficiency is low due to the slow growth of the plant, tubers and roots. In addition, due to the warm and humid environment, reproduction is limited to summer season. Due to the traditional reproductive problems in this plant, tissue culture or microbial culture is the best way to replicate rapidly and to achieve a large number of plants with the same genetic structure, as well as the elimination of diseases in the short term and reducing the costs. The aim of this study was to compare different microorganisms in terms of calogenesis and regeneration, as well as to determine the optimum culture medium for Zamiifolia tissue culture.
Materials and Methods: In this study, the explants prepared for the first experiment, including rhizome and petiole and the explants for the second experiment were the leaeaves and shoots. In the first experiment, rhizome and petiole were cultured in three replications in ½ MS medium containing BA (0, 2, 4 mg / L) and 2,4-D (0,1,2 mg / L) in combination with vitamins, 30 g/l sucrose, 5 g / L agar and adjusted to 5.8 PH. The cultivars were cultured for the callus induction under temperature of 27-25 ° C and light conditions of 16 hours light and 8 hours darkness. After 5 weeks, the percentage of callus and fresh callus weight were measured. The callus generated from rhizome and petiole in three replicates on ½ MS medium containing BA (0, 1, 2 mg / L) and 2,4-D (0, 0.5, 1 mg / l) for shoots and after the observation of branches and leaf buds were grafted on to ½ MS medium containing BA (0, 1, 2 mg / L) and NAA (0, 0.5, 1 mg / L) for rooting. Traits such as time to shoot elongation were recorded at regeneration stage, and after 5 weeks, shoot length and the number of leaves were measured. The time to rootstock was also recorded. In the second experiment leaf and shoot explants were cultured in ½ MS medium containing BA (0, 2, 4 mg / l) and NAA (0, 0.5, 1 mg / L) in combination with vitamins, 30 g/l sucrose, 5 g / L agar and PH adjusted to 5.8. The cultivars were cultured for the callus induction under temperature 27-25 °C and light conditions of 16 hours light and 8 hours darkness. The time to reach the callus was recorded and after 5 weeks, the percentage of callus and fresh callus weight were measured. The calli generated from the leaves and shoots were cultures on ½ MS medium containing BA (0, 1, 2 m g / L) and NAA (0, 0.5, 1 mg / L) for shoots and after observation of branch and leaves were transplanted to the ½ MS medium containing BA (0, 1, 2 mg / L) and NAA (0, 1, 2 mg /L) for rooting. The traits such as time to shoot elongation were recorded at the regeneration stage, After 5 weeks, the shoot length and the number of leaves were measured.The time to rootstock was also recorded.
Results and Discussion: The results of the first experiment showed that the effect of the rhizome and petiole type on the callus formation was significant at 1% level. So that the rhizome showed greater ability to callogenesis. The results of the second experiment showed that the effect of the type leaf and shoot on the callus formation was significant at 1% level. So that the leaf showed greater ability to callogenesis. The highest percentage of callosing (94.5%), the shortest time to reach the callus (14 days) and the highest callus weight (1.1 g) in culture medium with 2 mg / l BA and 1 mg / l hormones NAA was observed in leaf samples from the second experiment. The best treatment in the shoot elongation stage, which included the shortest time to shoot formation (10.5 days), the longest shoot length (4.10 cm), and the highest leaf number (8 leaves) in the leaf extract with hormonal concentrations of 2 mg / 0 mg / L NAA was observed from the second experiment. In the rooting stage, the best treatment for petiole extracts with hormonal concentrations was 1 mg / l BA and 0.5 mg / l NAA with 14 days to rooting from the first experiment.
Conclusion: In this study, explants and various concentrations of growth regulators had significant effect on the response to callus induction in Zamiifolia. In the first experiment, the rhizome and in the second experiment the leaf showed a better reaction to callus induction. According to this research, it can be suggested that the treatments applied in both experiments should be applied on all four leaves, petiole, rhizome and shoot samples, and the best culture type and the best culture medium for the cultivation of Zamiifolia plant tissue should be determined in subsequent studies.
Maedeh Aghdaei; Seyyed Hossein Nemati; Leila Samiei; Ahmad Sharifi
Abstract
Introduction: Pepino (Solanum muricatum Aiton) is a diploid herbaceous plant belongs to the Solanaceae family, which is growing in subtropical zone, originates from Andes in South America. It is commercially grown for its fruit, which is appreciated not only for food but also for its appearance, in South ...
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Introduction: Pepino (Solanum muricatum Aiton) is a diploid herbaceous plant belongs to the Solanaceae family, which is growing in subtropical zone, originates from Andes in South America. It is commercially grown for its fruit, which is appreciated not only for food but also for its appearance, in South American countries, including Bolivia, Colombia, Ecuador and Peru, as well as in countries such as New Zealand and Australia. Pepino is propagated by seed, cutting, and tissue culture methods. Most pepino cultivars are sexually fertile and produce viable seeds, but their seeds have poor germination and high level of heterozygosis causing to highly variable plants. Both mentioned negative aspects have limited the mass production of this plant through seed. In this case, stem cutting is used as the most common way of propagating pepino led to transmission of viral diseases and increasing propagation costs as two main limiting factors of pepino propagation. So, micropropagation systems are a promising tool to produce disease-free clonal plant material with low costs. Therefore, the present study was aimed to assess the effect of different media and plant growth regulators on micropropagation traits of pepino.
Materials and Methods: Three separate experiments were carried out in institute of plant sciences of Ferdowsi University of Mashhad in 2016. Pepino seeds were bought from company of Plant World Seed, UK, were cultivated on MS medium. Grown plants were used as source of providing explants. Four mediums, including MS, ½ MS, SH and B5 were used to determine the best culture medium for shoot regeneration of pepino using single node explant. A factorial experiment was conducted based on a completely randomized design. Some growth properties such as number of shoots, shoot length, number of roots, root length, leaf number and leaf length were evaluated after two and four weeks. In proliferation experiment, MS medium was compared with MS supplemented with different concentrations of BA (0.5, 1 and 2 mg L-1) and Kin (0.5, 1 and 2 mg L-1) applied as combined treatments, and also BA used alone at concentrations of 2, 4 and 6 mg L-1 that was conducted based on a completely randomized design. For rooting of explants, an experiment was conducted based on a completely randomized design containing of two concentrations of IBA (at 0.3 and 0.6 mg L-1) and three concentrations of NAA (at 0.3, 0.6 and 0.9 mg L-1) in MS medium. Some growth properties including root number and length, root density and root quality were evaluated after four weeks
Results and Discussion: Results indicated that micropropagation rate of pepino was affected by culture medium type. The highest shoot length, number of root, root length and leaf number were obtained in MS medium, although statistically there was no significant difference between MS and ½ MS media. The highest number of shoots and leaf length were observed in MS medium, which led to a significant difference with other media (½ MS, SH and B5). Overall, Based on obtained results MS medium was the best culture medium for micropropagation of pepino using single node. In the proliferation experiment, the highest shoot and leaf number and plant color were obtained with using 2 mg L-1 BA + 1 mg L-1 Kin, whereas the highest shoot length and leaf length were observed in the 1 mg L-1 BA + 2 mg L-1 Kin and 1 mg L-1 BA+1 mg L-1 Kin treatments, respectively. Increasing in concentration of BA up to 2 mg L-1 in combination with Kin had a positive effect on shoot proliferation, while applying BA at concentration 2, 4 and 6 mg L-1 alone led to decrease in proliferation. Results obtained from rooting experiment showed that the highest root number, root density and root quality were obtained using IBA at the concentration of 0.6 mg L-1, whereas the highest root length was observed by applying IBA at concentration of 0.3 mg L-1, which led to a significant difference with other treatments. Furthermore, results indicated that the effect of IBA on rooting of pepino microshoots was more than NAA.
Conclusion: Generally, the best results were obtained by MS medium, 2 mg L-1 BA with 1 mg L-1 Kin for shoot proliferation, and IBA at concentration of 0.6 mg L-1 for the rooting of pepino nodal segments.
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 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.
Maryam Aflaki Jalali; Abdollah Hatamzadeh; Hassan Bahrami Sirmandi
Abstract
Introduction: Tissue culture is an effective technique for mass propagation of walnut that has many advantages. Plants were obtainedby in vitro techniques in comparison with in vivotechniques areableto producet fruitearlier. However one of the major problems in walnut micropropagation is the difficulty ...
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Introduction: Tissue culture is an effective technique for mass propagation of walnut that has many advantages. Plants were obtainedby in vitro techniques in comparison with in vivotechniques areableto producet fruitearlier. However one of the major problems in walnut micropropagation is the difficulty of rooting. Auxin protection against auxin-oxidase system can make a major contribution to rooting. Among all the compounds that can play the synergistic role with auxin, they will probably have the ability of auxin protection against enzymes. In this experiment, the effect of lignosulfunate on rooting of micropropagated walnut was investigated for the first time.
Materials and Methods: In this experiment, Hartley cultivar of walnut was used. At first, explants were washed under running water for 1 hour then explants were placed in 70% alcohol for 1 minute and after that in 10% bleach for 10 minutes. After sterilization, under laminar air flow hood, explants were washed three times with distilled water and werecultured on Driverand Kuniyuki, 1984(DKW) medium supplemented with 2.2 g l-1phytagel, 2 mg l-1 BA, 0.01 mg l-1 IBA and 30 gl-1 sucrose (establishment stage). In multiplication stage, plantlets were subcultured every 25 days. All of the plantlets were placed in jars and were kept inside a growth chamber in photoperiod of 16 hours of light. All the multiplicated shoots were used as explants for the trails. Twodifferenttestswere usedto induceroot in explants. At the first trial, explants were transferred to induction medium containing IBA (3, 5, 7 and 10 mg l-1) and treatmentswere placedin thedark for 3, 5and7 days. Treatments related to theconcentrations of5and7 mg l-1IBAand7 daysof darknesshadthe highestpercentage ofrooting. In the next experiment, thecombination ofthree levels oflignosulfunate (1, 2 and 3 g l-1), and two concentration of 5and7mg l-1IBAwere used. Treatmentswere placedindarknessfor 7 days. After root induction,shootlets were transferred to root development medium. Rootdevelopmentmedium includesa quarteroftheDKWand vermiculite.
Results and Discussion:The aim of the first trial was to determine the concentration of IBA which produced the highest percentage of rooting. Among all the auxins, it was shown in other experiments that IBA has the best results in rooting of walnut. Due to this, we chose IBA as root induction hormone. With increasing of IBA concentrations and the induction period, rooting increased. Because the higher amount of exogenous auxin will induce the higher amount of endogenous auxin (IAAsp). However, rooting increased to a certain level and then began to decrease. With increasing concentrations above 10 mgl-1 IBA rooting reduced and formation of callus in the shoot end increased which is not good for rooting because callus would not let the cells form roots. It seems that accumulation of IAAsp induces self-productive cells in root area to grow and duplicate abnormally and maybe root formation stops because of this accumulation and also because of the inadequacy of the IAAsp to transfer to neighboring cells. Also with increasing concentration, defoliation and wilting happens. A lower concentration of IBA (about3 mgl-1) caused loss of rooting. The highest percentage of rooting for the first treatment with IBA was with 5 mgl-1 IBA and 7 days of darkness and 7 mgl-1 IBA and 7 days of darkness. The treatments were placed in darkness due to degradation of auxin under light condition. The induction time was related to auxin concentration. If the auxin concentration is less, the exposure time in the dark will be more. Root induction in the dark had better results than induction in light. The capacity of rooting in walnut is related to the amount of endogenous (IAAsp) and exogenous auxin. The amount of endogenous auxin is completely related to the cultivar and thatis why some cultivars respond really well to the amount of exogenous hormones in rooting stage. Exogenous auxin induces the production of endogenous auxin (IAAsp). These two concentrations were chosen for next treatment with lignosulfunate. Rootingratedecreaseswith increasinglignosulfunate. However, the highest root induction among all the treatments wasachieved on medium containing 1 g l-1lignosulfunate. The reason of transferring all explants after root induction to root development medium was changing the hormone and salts concentrations. At this stage, the ¼ DKW was used as a medium. This is due to the reduction of salts, root induction and rooting accelerate.
Conclusions: In this study, the effect of lignosulfunate (auxin synergist) on rooting stage of Hartley cultivar of walnut was investigated. For this goal, two trials were done. The first trial was to determine the best concentration of IBA for rooting. Two concentrations were chosen and another trial was the effect of the combination of lignosulfunate with IBA on rooting. For the first time in this study, we showed that lignosulfunatecan improve rooting of walnut.
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
Esmaeil Chamani; Mobina 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, ...
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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.
Askar Ghani; Ali Tehranifar; Valiollah Ghasemi; Samira Hatefi
Abstract
Pseudohandelia Tzvel. belongs to the Asteraceae family with only one species. It is endemic to Khorasan Province in Iran. Having partially big and beautiful inflorescences make it a proper choice in urban green space especially in case of mass culture. In this research, to investigate possibility of ...
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Pseudohandelia Tzvel. belongs to the Asteraceae family with only one species. It is endemic to Khorasan Province in Iran. Having partially big and beautiful inflorescences make it a proper choice in urban green space especially in case of mass culture. In this research, to investigate possibility of micro propagation of this plant two separate experiments were designed. To obtain non contaminated explants, seeds were cultured in MS media in February 2009. After about two months a few healthy plants as stock plants for use in later stages produced through subculture. In the first stage, to evaluate amount of callus production, a factorial experiment based on a randomized complete design with two factors conducted. The first factor was 12 different hormone levels (including BAP, IAA, KN and 2,4-D with different concentrations) and second factor was explants type (including leaf, apical and lateral buds as a explants) making 36 treatments in total. In the second stage to induce stem on produced callus a randomized complete design with 34 treatments (including mentioned hormonal treatments, base MS media and different explants) was used. At the end the callus with stems were transplanted to a MS media containing IBA for root production. The results of the first experiment showed callusogenensis response to above hormonal treatments was seen in this plant with the exception of 2,4-D. Significant difference among the means of hormonal treatments for fresh weight of induced callus was observed. The result of the second experiment showed limited treatment could produce stem and most of treatments just produce callus. Also those samples that produced stem, a few could produce root in MS media.
Ahmad Reza Bolandi; Hassan Hamidi; Roya Beidokhti
Abstract
The experiment was conducted to study the effect of photoperiod and hormones on production of microtubers for two potato cultivars (Sante and Savalan), which were clean and pathogens free. This experiment was performed as a factorial experiment based on randomized design with three replications. The ...
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The experiment was conducted to study the effect of photoperiod and hormones on production of microtubers for two potato cultivars (Sante and Savalan), which were clean and pathogens free. This experiment was performed as a factorial experiment based on randomized design with three replications. The results showed that cultivar, hormone and photoperiod would effect on all studied traits. Sante cultivar indicates more efficiency for all measured traits than in Savalan cultivar. In this experiment by using the combination of two hormones 2, 4-D and BAP would increase number, diameter and weight of microtubers. The means comparison of photoperiod showed that highest efficiency for all traits as treatment of plant for 8 hours in darkness plus 16 hours in light plus utter darkness (P3). In this research, the highest number of microtuber was related to cultivar of Sante (9.47) is gained with hormone 2, 4-D, as well as photoperiod P3 which this superiority for Savalan cultivar is gained by using the combination of two hormones and P3 photoperiod.
Shadi Mohamadi-Nejad; M. Gholami; Mahmood Esna-Ashari
Abstract
This research was conducted to study the factors affecting establishment, growth and shoot proliferation of Persian walnut, genotype Z60. Explant establishment was studied through a factorial experiment based on a completely randomized design with two factors including culture media (DKW and WPM) and ...
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This research was conducted to study the factors affecting establishment, growth and shoot proliferation of Persian walnut, genotype Z60. Explant establishment was studied through a factorial experiment based on a completely randomized design with two factors including culture media (DKW and WPM) and BA concentration (0 (control), 0.5 and 1.0 µM). Single-node explants were prepared from the young branches of Z60 trees in the middle of May. Although the effect of two culture media on the growth characteristics had no significant difference, but mean comparison of the data with this regard showed that DKW medium performed better than WPM. No significant difference was observed between the two BA concentrations, but significant difference was between two concentrations (0.5 and 1.0 µM) with 0 (control). In shoot proliferation experiment, two cytokine hormones (BA and Kinetin) in three concentrations (4.4, 6.6 and 8.8 µM) were studied in DKW medium. The data obtained from this part of experiments were analyzed through unbalanced completely randomized design. The effect of different hormonal treatments and their concentrations on Z60 genotype growth characteristics showed that 8.8 µM BA was more effective in compared to the other two treatments. However, no significant difference was observed between the three concentrations with this respect.
