mehdi mohebodini; Zahra Azimzadeh; Esmail Chamani; Malihe Erfani
Abstract
Introduction: Lily (L. ledebourii) is the rarestspeciesof thegenusLilium, and grows in Caucasus region. Iranis one of the important distribution areas of this endangered species. It is important as an ornamental plant due to its large and attractive white flowers that are equal to those of commercial ...
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Introduction: Lily (L. ledebourii) is the rarestspeciesof thegenusLilium, and grows in Caucasus region. Iranis one of the important distribution areas of this endangered species. It is important as an ornamental plant due to its large and attractive white flowers that are equal to those of commercial lilies in terms of beauty.The two main constraints on growing this plant are a low multiplication rate and the high cost of bulb production. Five to ten flowers commonly appear on each plant, even specimens with up to 15 flowers have been observed. Plant tissue culture techniques are widely used in plant propagation and using these methods can effectively provide micro-propagation of this plant in large scale. High percentage ofregeneration is necessary for plant protection, using in the breeding programs and gene transfer to this plant. Therefore, the effect of plant growth regulators and abiotic stress (ultrasound) werestudied on the bulblet production and root induction of Lilium ledebourii.
Materials and Methods: The experiment was factorial based on completely randomized design with four replicattions and was carried out in tissue culture lab of University of MohagheghArdabili in 2015. For this purpose, segmentsof scale explant was treated with ultrasound and cultured on MS medium supplemented with different concentrations of NAA and BA alone and/or in combination with each other. In this experiment, different concentrations of NAA (0, 0.01, 0.1 and 1 mgl-1) and BA (0, 0.01, 0.1 and 1 mgl-1) and different Ultrasound exposure duration (0, 5, 10, 20 and 30 second) were studied. In order to remove possible contamination from the media, all media were autoclaved for 20 minutes at 121 °C. At the end of the experiment, the number of bulblet, root length, fresh weight of bulblet were recorded. The cultures were kept at 25±2°C under illumination with daylight fluorescent lamps (30 mol m-2s-1) at 16 h photoperiod. Data was subjected for analysis of variance and compare means using SPSS 16.
Results and Discussion: The results showed that ultrasound had negative effect onroot length, so that the highest root length was observed in explants without ultrasound treatment. Result also indicated that ultrasound had positive effect on bulblet production and root induction. A different effect of growth regulators was observed in similar media on the bulblet formation percentage. The 0.1 NAA concentration had a higher efficiency while increasing NAA insignificantly decreased bulblet induction. The highest total weight and number of bulblets obtained by 0.1 mgl-1 NAA. Concentrations of NAA increased rooting percentage. Different concentrations of NAA had also significant effects on some traits. So that, the highest weight of bulblets obtained by 0.01 and 0.1 mgl-1 BA and the highest number of roots obtained in control. Bulblet maximum mean weightwas in30 seconds ofultrasoundtreatment, which hada significantdifference with the control treatment (without ultrasoundtreatment). In the other hand, ultrasound increased the number and weight of bulblets.Mechanical stress and microstreaming by acoustic cavitation might be considered as the most possible cause of the various physiological effects of ultrasound on cells. The enhancement of V-ATPase transport and ATP hydrolysis activities seem to be an ultrasound-induced metabolic response of cells. High-intensity ultrasound is well known to be destructive to biological materials, disrupting the cell membranes and deactivating biological molecules such as enzymes and DNA. Low-intensity ultrasound, on the other hand, has shown a range of sub lethal biological effects that are of potential significance in biotechnology. There are several processes that take place in the presence of cells or enzymes activated by ultrasonic waves. High-energy ultrasonic waves break the cells and denature the enzymes. Low-energy ultrasound can modify cellular metabolisms or facilitate the uptake of nutrient, and make them easily through the cellular walls and membranes. In the case of enzymes, the increase in the mass transfer rate of the reagents to the active site seems to be a most important factor.
Conclusions: The results showedthatthebulblet production at first stages and a little root formation in tissue culture is useful for fast bulblet inductionandthenrooting. Finally, it seems that ultrasound in combination with plant growth regulators have the potential to produce the highest average number of bulblets in the scale explant.
zahra shabani; Bahram Abedi; Abrahim Gangimoghadam; Ali Tehranifar
Abstract
Introduction: This study was conducted aimed to consider the effects of culture medium and the concentration of growth regulators on proliferation, rooting and the acclimatization of Myrobalan29C. This study was performed as a factorial experiment in a completely randomized design (CRD) with four replications ...
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Introduction: This study was conducted aimed to consider the effects of culture medium and the concentration of growth regulators on proliferation, rooting and the acclimatization of Myrobalan29C. This study was performed as a factorial experiment in a completely randomized design (CRD) with four replications where each plot contained five explants. Given the role of trees rootstock of growth rate vegetative in the early maturity٫ yield and disease resistance will be suitable rootstock that has an important role in program garden management. In total rootstock of fruit trees have be propagation sexual and asexual methods, however٫ given that in sexual reproduction the resulting dispersion characteristics and the resulting seedlings changed by genetics, it is tried for decades to asexual propagation methods in specially tissue culture methods healthy rootstock for mass propagation and used the development of orchards. Seedling rootstock Myrobalan had been used in the past. The rootstock can be positive features cited the ease of access, to be cheap, good yields after maturity. In study, the effects of NAA and BAP on proliferation of Gisala5 rootstock showed the most shoot treated in mediacontaining 1mg/l BAP. Investigated Chinese plum in vitro micropropagation showed that 1/2 MS media had the highest percentage rooting, and acclimatization rooted plantlets to greenhouse conditions was using the system miss successfully. Due to the importance and essential to achieve an efficient protocol for the mass propagation of Myrobalan 29C in Iran, this study was conducted with the main purpose of evaluating the most suitable media culture and plant growth regulators in micropropagation of Myrobalan 29C.
Material and methods: The explants were collected from shoots of Myrobalan 29C rootstock maintained in the experimental greenhouse of Khorasan Razavi Agricultural and Natural Resources Research Centre (of Mashhad, Iran), on June 25, 2013. The explants were washed by water and dishwashing liquid to removed surface contamination. Then they were divided to some parts containing one bud and were sterilized with ethanol 70% 1 min and sodium hypochlorite 10% at 10 min. Proliferation was performed in two kind of culture medium (MS and DKW) that supplemented with plant growth regulators BAP (0, 1, 2, 3, 4 mg l-1). The rootstocks of in this step, after subculture three (21 days between each subculture), the numbers, the length and quality of the shoots (explants strong growth, with no signs of vitrification, necrosis of leaf are yellowing terminal meristem), b– less than 15% have the symptoms of vitrification, necrosis of leaf are yellowing terminal meristem, and c– explant weak, 15-30% have the symptoms of vitrification, necrosis of leaf are yellowing Terminal meristem) were measured. This stage was carried with four replications and each replicates with five samples. Two culture media (MS and DKW) were used for rooting, which supplemented with indole-3-butyric acid (IBA) at four levels (0, 1, 2, 3 mg l-1). This stage was carried with four replications and each replicates with three samples. After being rooted explants, the best cultured media and combination of rooting growth regulators number and root length, leaf number and stem length and quality of explants were recorded. Acclimatization used in substrate including Coco Peat - Perlite 3:0/5 V, Coco Peat 100%V٫ Coco Peat - Perlite 0.5:3V, Perlite100%V and Coco Peat - Perlite1:1V).
Results and Discussions: Results showed that the 10% sodium hypochlorite for 10 min, with 4% decay was the best treatment for sterilization. The results showed that the proliferation average was 6/16 in MS medium with 2 mgl-1 BAP and the most percent of rooting and root length were about 100% and 2/51 cm in MS medium with 2mgl-1 of IBA, respectively. The acclimatization of plantlets to greenhouse conditions was successful. The highest rate of plantlets survival (about 70%) was obtained from substrate Cocopit and Prlit (1; 1 V). In the present study, explants year compared annual explant least contamination had enjoyed. It seems that the young explants, smooth the surface of the skin, having the least amount of crack depend on the type of surface depressions explants and the crack and lower depressions the surface explants increase surface contact area disinfectants and also improve its impact. In this study found that the type of medium a significant impacted on the health of plants and so the proliferation of explants was successful. Usually root production in plants under the influence of synthesis, metabolism, and transport is auxin signaling pathways. Therefore acclimatization directly affected the rooting of plants that had high quality and the best rate of induction.
Conclusions: The results of this research showed that we can duplicate Myrobalan29c rootstock by in vitro method. According in this research, MS media including BAP and IBA plant growth regulators are the most suitable for micro propagation.
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; K. 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
G. Chenarani; Mahmoud Shour; Ali Tehranifar; S.H. Nemati; Gholam Hossein Davarynejad
Abstract
CO2 enrichment in greenhouse is a suitable way which reduces production time, better growth vigor and also higher plant quality. The main aim of this study was to find out the effects of artificial CO2 enrichment under different light levels on rooting of the ornamental foliage Codiaeum variegatum. The ...
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CO2 enrichment in greenhouse is a suitable way which reduces production time, better growth vigor and also higher plant quality. The main aim of this study was to find out the effects of artificial CO2 enrichment under different light levels on rooting of the ornamental foliage Codiaeum variegatum. The experiment was planned as a split plot based on a completely randomized design. CO2 was considered as the main plots (380 as control, 750 and 1050 ppm) and these light intensities as the sub plots (10000 as control, 12000 and 14000 Lux) were used. Results showed a significant increase on measured traits with elevating levels of CO2 and light. Highest measured values of different traits were observed at 12000 Lux light intensity and 750 ppm CO2 enrichment. Light intensity × CO2 interaction had a significant effect on leaf length, leaf number, root quality, root volume, root length (P≤0.01) and rooting percentage at (P≤0.05). Spad chlorophyll unit was not noticeably significant. Rooting and plant growth generally raised along with both light intensity and CO2 elevation.
M. Shoor; M. Behzadi; M. Goldani
Abstract
Carbon dioxide (CO2) concentration of the global atmosphere has increased during the last decades. Increasing global atmospheric CO2 concentrations are expected to influence on plants. Coleus is an ornamental plants, that is due to its attractive foliage are considered. In order to evolution of high ...
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Carbon dioxide (CO2) concentration of the global atmosphere has increased during the last decades. Increasing global atmospheric CO2 concentrations are expected to influence on plants. Coleus is an ornamental plants, that is due to its attractive foliage are considered. In order to evolution of high CO2 concentration effects on rooting. morphological and anatomical traits in two coleus spices (C. scutellarioides and C. blumei) a factorial experiment based on completely randomized design with 3 replications and 6 treatments were conducted at the greenhouses of Ferdowsi University of Mashhad at 2010. Treatments were two coleus spices and 3 concentration, of CO2 380(as a control), 700 and 1050 ppm. Leaf cuttings plants were placed under increasing CO2 concentration during of 30 days. The number of leaves, stem diameter wet weigh, dry weight, length of stoma, width of stoma, size of stoma and stomatal density were measured. Results indicated that increasing CO2 concentration from 380 to 1050 ppm leading to increase of 25.3, 32.4, 20.5 and 75 percentage in number of leaves, stem diameter, wet weigh and dry weight respectively. Also these results showed that with increase of CO2 concentration decreased length of stoma, width of stoma and size of stoma, but increased of stomatal density. The highest mean density of stomatal to 30.7 per mm2, which was related to the interaction of carbon dioxide concentration of 1050 ppm and C. is scutellarioides than other treatments showed significant differences.