Tissue culture and Micropropagation
Zeinab Chaghakaboodi; Mahdi Kakaei; Alireza Zebarjadi; danial kahrizi
Abstract
Rapeseed (Brassica napus) is recognized as one of the most important oilseed crops worldwide and its development of cultivation has received attention due to the importance of importing oil. The current study aimed to investigate the effect of drought stress on some Rapeseed genotypes under tissue culture ...
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Rapeseed (Brassica napus) is recognized as one of the most important oilseed crops worldwide and its development of cultivation has received attention due to the importance of importing oil. The current study aimed to investigate the effect of drought stress on some Rapeseed genotypes under tissue culture and field conditions and to identify stable genotypes in the field. The possible responses of 14 different canola genotypes to Callus induction resulting from Hypocotyl cultivation and evaluation their drought tolerance were studied using Polyethylene Glycol 6000 (PEG 6000) at five different levels, including zero (as control), 10%, 20%, 30%, and 40% PEG concentrations based on a completely randomized design (CRD) with three replications. Measured traits included relative growth rate, growth rate, relative water content, and proline content of the Callus. Furthermore, in the field sector, the genotypes were investigated in four environments (two consecutive years in 2016-2018 under rainfed and irrigated conditions) based on randomized complete block design with three replications. According to the Callus culture results, the assessed traits, except the Proline content of Callus, decreased with increasing stress level. In laboratory conditions, genotype number seven (Dante) was introduced as the superior genotype. The results of Additive Main effects and Multiplicative Interaction (AMMI) analysis showed the significance of both additive effects of genotype and environment and the multiplicative effect of genotype × environment interaction. The results of cumulative additive effects (decomposition of variance) and multiplicative interaction effects (decomposition into principal components) showed that the first two components explained 53.02 and 33.65% of the variance of the interaction effect for oil yield. Dante and SLM-046 genotypes were introduced as stable genotypes.
Tissue culture and Micropropagation
Eshaghali Bayati; Masoud Gomarian; Hossein Mirzaie Nodoushan; Mahdi Changizi; Shahab Khaghani
Abstract
In a laboratory experiment conducted in Kabudarahang, Hamadan, a promising potato genotype was selected through somaclonal variation from Agria commercial cultivar, based on the callus size on growth media, shoot and root regeneration. A field experimental design with three replications was carried out ...
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In a laboratory experiment conducted in Kabudarahang, Hamadan, a promising potato genotype was selected through somaclonal variation from Agria commercial cultivar, based on the callus size on growth media, shoot and root regeneration. A field experimental design with three replications was carried out to assess the new genotype along with its parental commercial cultivar for several morphological traits. Results revealed that total dry matter per plant (13.5%), total number of tubers (12.3%), tuber weight per plant (8.6%), and stolon length (4%) were significantly higher than in the new genotype. Also, days to 50% flowering (11.4%) and maturity date (7.6%) were significantly less in comparison with the parental cultivar. Several nutritional traits were also studied by which more antioxidant activity was observed on the new genotype (35.6%) than Agria cultivar (25.3%). Furthermore, the new genotype had a lower nitrite (81.2%) and higher iron (20.7%) and starch content (21.2%) comparing with Agria cultivar. Therefore, it seems that the new genotype has an acceptable potential for cultivation and replacement with existing cultivars.
Tissue culture and Micropropagation
Roghieh Sharifpour; Mohammadali Ali Aazami; Mohammad Bagher Hassanpouraghdam
Abstract
Stevia rebaudiana Bertoni is a perennial shrub from the Asteraceae; native of Brazil and Paraguai. This Plant is containing sweet steviosides; rebaudioside and isostevol. This experiment was conducted to study the effects of diverse factors on induction and growth of hairy roots induced by Agrobacterium ...
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Stevia rebaudiana Bertoni is a perennial shrub from the Asteraceae; native of Brazil and Paraguai. This Plant is containing sweet steviosides; rebaudioside and isostevol. This experiment was conducted to study the effects of diverse factors on induction and growth of hairy roots induced by Agrobacterium rhizogenes. The seeds were surface sterilized and then cultured on solid MS medium under 250C and 16:8 h photoperiod. After germination and development of seedlings, the root, stem and leaf explant of three different ages (30, 60 and 90 days) were placed on NYA and LB suspension media. Agrobacterium rhizogenes strains (ATTCC15834, R1000 and C58) and acetosyringone were treated on the plant sample by floating and spray method for 72 hrs at 270C. The incubated explants were treated to remove any bacteria and were transferred to 1/2MS media containing 400 mgl-1 cefotaxime. To optimize the growing media for hairy root induction and growth, 1/2 MS medium was enriched with 3% sucrose and 0.2 mgl-1 Indole-3-butyric acid. The results revealed that all the Agrobacterium strains were effective on hairy root induction from root and leaf explant. The highest hairy root induction was belonged to NYA suspension medium co-cultured with 1/2 MS. Furthermore, for the roots proliferation, 1/2MS medium containing 1/5% sucrose was the selected one. Leaf clone (C2LA) stevia lines obtained from ATCC15834 strain had the highest growth.
Tissue culture and Micropropagation
Pegah Moradi Dezfouli; Mohammad Sedghi; Mehran Enayati Shariatpanahi; Bahram Alizade
Volume 7, Issue 18 , November 2017, , Pages 1-14
Abstract
In this research, Hayola F1 hybrids were used to produce rapeseed doubled haploid lines using microspore embryogenesis. To study general combining ability (GCA) of, the induced doubled haploid (DH) rapeseed lines, a top cross analysis was conducted using 28 doubled haploid lines and top cross parent ...
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In this research, Hayola F1 hybrids were used to produce rapeseed doubled haploid lines using microspore embryogenesis. To study general combining ability (GCA) of, the induced doubled haploid (DH) rapeseed lines, a top cross analysis was conducted using 28 doubled haploid lines and top cross parent of Hayola 420. Produced hybrids of doubled haploid lines × Hayola 420 were sown in research farm in 2015 growing season. Plant height, number of pods per branch and sub branches, number of seeds per pod, pod length, number of sub branches, length of main branch, 1000-seeds weight, single plant yield, number of days to flowering, number of days to seeding, number of days to physiological maturity, and oil yield were recorded in all top cross progeny to investigate GCA of DH lines. Results of analysis of variance showed a significant difference among all top cross hybrids for all investigated traits at 1% probability level. Based on means comparison analysis using multiple range Duncan test at 1% probability level, top cross hybrids of DH1, DH8, DH10, DH11, DH13, and DH21 were more differ than other top cross hybrids for all investigated characteristics. The highest mean of seed yield and oil yield was related to the top cross progeny of DH21 × Hayola 420. Results of top cross analysis showed that the highest positive and significant GCAs for single plant seed yield, number of pods per plant, and 1000-seeds weight were corresponded to DH1, DH10, and DH21, therefore these three DH lines can be used as elite parental lines in future breeding programs of rapeseed.
Tissue culture and Micropropagation
Ebrahym Beiramizadeh; Roghayeh Zarei; Zohreh Hajibarat; Zahara Hajibarat; Abbas Saeidi
Volume 7, Issue 18 , November 2017, , Pages 93-102
Abstract
In vitro propagation has proven a suitable method for mass multiplication of uniform and diseases-free plants and acts as a new tool for modern breeding through genetic manipulation. This experiment was conducted to study the in vitro effects of growth regulators on proliferation and rooting ability ...
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In vitro propagation has proven a suitable method for mass multiplication of uniform and diseases-free plants and acts as a new tool for modern breeding through genetic manipulation. This experiment was conducted to study the in vitro effects of growth regulators on proliferation and rooting ability of Rosa canina. Auxiliary buds were used as explant in this experiment. The stem formation was tested using completely randomized factorial design. MS medium for shoot proliferation was supplemented with various concentrations of BA (0.5, 1, 1.5, 2 mgl-1) and Kin (0, 0.5 mgl-1). Analysis of variance showed that shoot number and shoot height were highly significant at 1% level. The most shoot proliferation was observed at 1 mgl-1 BA with 0.5 mgl-1 Kin. Maximum plantlet length was obtained with 0.5mgl-1 BA and 0.5mgl-1 Kin in combination. Number of roots, ratio of root number to root length and root length showed statistically significant difference in response to different root induction treatments. Furthermore, best root regeneration was obtained at 1.2 mgl-1 IBA in R. canina. All hardened plantlets were transferred to commercial rose greenhouse.
Tissue culture and Micropropagation
Peyman Sharifi; Ahmad Moieni
Volume 6, Issue 13 , May 2016, , Pages 13-26
Abstract
The effect of some factors on embryogenesis from isolated microspore and plantlet regeneration from microspore-derived embryos was studied separately in Brassica napus. Experiments carried out in factorial based on completely randomized design. The first factor in all of the experiments was cultivars ...
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The effect of some factors on embryogenesis from isolated microspore and plantlet regeneration from microspore-derived embryos was studied separately in Brassica napus. Experiments carried out in factorial based on completely randomized design. The first factor in all of the experiments was cultivars contained Global, Option and PF7045/91. The microspores were isolated from 3-4 mm buds and cultured on NLN-13 medium. Cultures incubated at 30˚C and darkness for 14 days, and then transferred to shaker in the growth chamber at 25˚C. In regeneration experiment, embryos with 20-25 days old were transferred to B5 medium and nearly 20-25 days after transferring embryos, normal regenerated plantlets, abnormal regenerated plantlets, rooted embryos and non differentiated embryos were counted. In the first embryogenesis experiments, the effects of medium volume, cultivar and interaction effects of two factors were significant. In Global cultivar, the highest values of embryos (695.5 per Petri) were observed in 12.5 ml medium volume. In the second embryogenesis experiment, the interaction between activated charcoal and cultivar on embryogenesis was significant. In the third embryogenesis experiment, the form of carbohydrate had a significant effect on embryo yield. In the first experiment of plantlet regeneration, GA3 had significant effect on normal regenerated plantlets, abnormal regenerated plantlets, rooted embryos and non differentiated embryos and the interaction between GA3 and cultivar was significant for normal regenerated plantlets. In the second experiment of plantlet regeneration, the interaction between gelling agent and cultivar was significant on normal regenerated plantlets.
Tissue culture and Micropropagation
Behzad Ahmadi; Rasoul Asghari Zakaria; Mehran Enayati Shariat Panahi; Naser Zare; Pejman Azadi
Volume 5, Issue 10 , September 2015, , Pages 17-29
Abstract
In this study, the effect of cold treatment (4 °C for 1 to 5 days) in combination with heat shock (30 °C for 1 to 10 days) and also colchicine treatment (25 to 100 mg/l for 24 to 72 h) were assessed on induction of sporophytical divisions in isolated microspore culture in two hybrid tomato cultivars ...
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In this study, the effect of cold treatment (4 °C for 1 to 5 days) in combination with heat shock (30 °C for 1 to 10 days) and also colchicine treatment (25 to 100 mg/l for 24 to 72 h) were assessed on induction of sporophytical divisions in isolated microspore culture in two hybrid tomato cultivars (‘Berlina’ and ‘Petoperide’). Microspore-derived structures with more than 10 nuclei were only observed in cv. ‘Berlina’ and in the cultures incubated for 1 or 2 days at 4 °C and then for 2 days at 30 °C. In addition, cold treatment for 1 or 2 days and then 2 days at 30 °C could efficiently induce formation of microspore-derived structures with 9-10 nuclei in both cultivars tested. No microspore with more than 5 nuclei was observed in the cultures treated at 30°C for 10 days. In the cv. ‘Berlina’, microspore-derived structures with 9-10 nuclei were detected when 25 mg/l colchicine was used for 48 h, while in cv. ‘Petoperide’, microspore-derived structures with more than 8 nuclei were not observed in all treatments tested. Globular embryos were only produced in two-layered culture medium when treated at 4°C for 2 and 5 days and then subjected to 30°C for 2 days. Microspore embryogenesis could be induced in tomato if appropriate duration of cold and heat treatment was selected.
Tissue culture and Micropropagation
Keyghobad Kaikavoosi; Altaf Hosseini Nadaf; Gholamreza Bakhshi khaniki
Volume 5, Issue 9 , June 2015, , Pages 29-38
Abstract
In most transformation studies it has been indicated that adding proline to the tissue culture medium can increase the callus induction frequency and reduce induction time. Adding proline to callus induction medium in this phase can affect the production of aromatic compounds in rice and if the goal ...
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In most transformation studies it has been indicated that adding proline to the tissue culture medium can increase the callus induction frequency and reduce induction time. Adding proline to callus induction medium in this phase can affect the production of aromatic compounds in rice and if the goal of exogenous gene transformation is increasing the rice aromatic associated secondary metabolites such as 2-acetyl-1-pyrroline (2AP), this amino acid can slipup the final results. Although, absence of amino acids such as proline can reduce callus induction percentage. In this research, callus induction from two indica rice varieties; Ambemohar 157 and Indrayani were optimized, using MS medium having various concentrations of 2,4-D, without using proline. The results revealed that 2.5 mgl-1 of 2,4-D for Ambemohar 157 and 4 mgl-1 for Indrayani can lead to better callus induction. These results indicated that absence of proline can be disregard by increasing of 2,4-D concentrations. Calluses obtained from the best hormone treatment were cultured on MS fortified with 0.01 mgl-1 NAA + (1, 2, 3, 4, 5) mgl-1 BAP for shoot regeneration. The highest percentage of regeneration was achieved on MS supplemented with 2 mgl-1and 3 mgl-1 for Ambemohar157 and Indrayani cultivar respectively. Proline contents in calli which were growth in MS medium supplemented with 500 mg/L of proline showed approximately 12 to 14 fold increase over the calli growth in non-proline added medium.
Tissue culture and Micropropagation
Ali Boosh; Ahmad Moeini; Hamid Dehghani; Zahra Movahedi
Volume 4, Issue 8 , March 2015, , Pages 47-55
Abstract
The plant bioreactors can be used for in vitro plant propagation and leading to a decrease in the costs of micropropagation. In this research, a simple bioreactor including a temporary immersion system was designed and made for Damask rose micropropagation. To optimize this system, the effects of the ...
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The plant bioreactors can be used for in vitro plant propagation and leading to a decrease in the costs of micropropagation. In this research, a simple bioreactor including a temporary immersion system was designed and made for Damask rose micropropagation. To optimize this system, the effects of the tube type, container size and different substrates were investigated in the independent experiments. Also, the temporary immersion system was compared with the solid and liquid media. Results showed that the highest number of the shoots in the temporary immersion system were obtained by the use of the white tube (11 shoots per explant), 600 ml glass container (10.36 shoots per explant) and without any substrate (10.76 shoots per explant). Also, temporary immersion system was significantly better than the solid and liquid media for shoot production (10.66, 2.56 and 5.66, respectively).
Tissue culture and Micropropagation
Maryam Tavakoli; Mehran Enayati Shariatpanahi
Volume 3, Issue 4 , September 2013, , Pages 61-68
Abstract
In this study, an experiment was designed for optimization of mutation application on microspore embryogenesis of canola, regeneration of embryos and producing haploid plants, from Hayola 401 and Hayola 402 hybrids using EMS mutagen treatments. EMS stocks were prepared for 0.1, 0.2 and 0.3 percent using ...
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In this study, an experiment was designed for optimization of mutation application on microspore embryogenesis of canola, regeneration of embryos and producing haploid plants, from Hayola 401 and Hayola 402 hybrids using EMS mutagen treatments. EMS stocks were prepared for 0.1, 0.2 and 0.3 percent using NLN13 culture medium and then microspores were cultured in NLN13 medium. Then, they were stressed by heat shock (30°C for 14 days). The results showed that in both hybrids by 0.1 % mutagen treatment for 90 minutes, the highest numbers of embryos achieved. So in the 90 minutes durations all of three applied concentrations from EMS and the control had significant differences and by increasing the concentration along this duration, fewer embryos were achieved. Afterwards, embryos were transferred to B5 medium containing gibberellic acid and incubated at 25°C and after full growth they were transferred to B5 medium without gibberellic acid. The results in both hybrids showed that the highest numbers of regenerates were produced when EMS (0.3% for 90 min) was applied to isolated microspores.
Tissue culture and Micropropagation
Hamed Ebrahimizadeh; Mahmoud Lotfi; Shiva Azizinia; Farangis Ghanavati
Volume 3, Issue 4 , September 2013, , Pages 99-108
Abstract
Production of haploid plants was studied in summer squash via induction of parthenogenic embryos. Female flowers were pollinated with anthers irradiated by different doses of gamma ray (25, 50, 75, 100 and 200 Gray) and induced embryos were rescued and cultured in specific medium. The results achieved ...
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Production of haploid plants was studied in summer squash via induction of parthenogenic embryos. Female flowers were pollinated with anthers irradiated by different doses of gamma ray (25, 50, 75, 100 and 200 Gray) and induced embryos were rescued and cultured in specific medium. The results achieved showed that the highest number of embryos was obtained in 50 and 75 Gray doses (82%), however, no embryo was produced at 200 Gray. Gamma ray doses and embryo stage had significant effect on frequency of embryo and plant regeneration. Highest embryo regeneration and haploid formation (27 plant regeneration and 11 haploids formation, respectively) were recorded at 50 Gray. All amorphous embryos produced only diploid plant, though 29.17, 33.33, 57.14, 66.67, 100 and 100 percent of plants derived from cotyledon, heart, torpedo, arrow-tip, torpedo-tip and globular embryos respectively, were haploid. Based on this study, out of the 7744 extracted seeds, 127 embryos and 44 plants were regenerated; among those 17 plants were identified as haploid plants using chloroplast counting, Flow-Cytometry technique and morphologic traits evaluations.
Tissue culture and Micropropagation
Abdolreza Bagheri; Fereshteh Moshiri; Sara Khosravinia
Volume 3, Issue 5 , February 2013, , Pages 53-61
Abstract
Black zira is a perennial herb with highly valuable impact in the pharmaceutical industries. To overcome the existing limitations of the field, achieving efficient regeneration methods provides the possibility of genetic manipulation and improvement of traits in this plant. Our objective in this research ...
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Black zira is a perennial herb with highly valuable impact in the pharmaceutical industries. To overcome the existing limitations of the field, achieving efficient regeneration methods provides the possibility of genetic manipulation and improvement of traits in this plant. Our objective in this research work is to develop an efficient regeneration protocol of bleack zira being used in genetic improvement programs. After optimizing the conditions for sterilization and seed germination, sterile cotyledonary seedlings were obtained on MS basal medium. We investigated efficient callus induction, rooting and shoot regeneration methods on Murashige and Skoog’s medium containing different concentrations of growth regulators including BAP, NAA and IAA in a factorial experiment based on completely randomized design with 3 replications. A significant increase in callus, rooting and regeneration from rootlet explants was observed. We noted that BAP and NAA were more capable for callus initiation and long-persistent development. It is shown that the most frequency and dry weight of callus was obtained on medium MS supplemented with 0.5 or 1mg/l BAP and NAA. Often frequency of rooting was influenced by NAA growth regulator. Also BAP and IAA in single stage regeneration in this study were found to be effective so that the addition of IAA to the culture medium caused reduction in callus development and showed direct shoot regeneration from cotyledon node, hypocotyl and rootlet explants.
Tissue culture and Micropropagation
M Dehestani Ardakani; M Kafi; M Enayati Shariatpanahi; M Jafarkhani-Kermani; M.R Fattahi Moghadam; M Oroojloo
Volume 2, Issue 3 , January 2013, , Pages 73-83
Abstract
One of the most commonly used methods to produce doubled haploid plants is microspore culture. In this research work, the isolated microspore culture system in two rose cultivars i.e. Apollo and Amarosa was investigated. Important factors including isolation media (AB, B), AT3 induction medium with different ...
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One of the most commonly used methods to produce doubled haploid plants is microspore culture. In this research work, the isolated microspore culture system in two rose cultivars i.e. Apollo and Amarosa was investigated. Important factors including isolation media (AB, B), AT3 induction medium with different carbohydrate sources (sucrose, maltose or glucose) and amino acid source (lactalbumin hydrolysate) were studied. Carbon starvation and temperature (heat and cold) treatments as two important stresses alone or in combination with each other for various periods were evaluated on the induction of symmetrical (sporophytic) divisions. A mixture of different developmental stages of microspores was used to initiate the cultures but the majority of them were at late uni-cellular stage. For eliminating bacterial or fungal contaminants, buds were surface-sterilized by immersion in 70% ethanol for 15, 30, 60 Sec. or 3.5% (w/v) sodium hypochlorite solution for 5, 10, 15 min prior to microspore isolation. The best sterilization procedure was observed when microspores were sterilized with sodium hypochlorite (%3.5) for 10 minutes. Two isolation media did not show a significant difference on the viability of microspores. Among induction media tested, in cv. Amarosa, the highest viability was observed in AT3 medium supplemented by glucose. Induction media in Apollo cultivar did not show a significant difference on viability of microspores. Combination of starvation (B medium) and cold (4°C) treatments for 3 days induced formation of pro-embryos in cv. Amarosa. Present investigation reports a protocol for induction of embryogenic developement in rose (Rosa hybrida) microspores.
Tissue culture and Micropropagation
Bahareh Tayefe-Afshari; Mehran Enayati Shariat Panahi; Mojtaba Vahab-Zadeh
Volume 1, Issue 1 , March 2012, , Pages 13-21
Abstract
The objective of this study was to improve induction of embryogenesis in bread wheat microspores. In this study, some F1 hybrids i.e M85-6 × 90, M85-8 × 90, mv17 × shiroudi, mv17 × kavir and kavir × bam were used. Microspores were cultured in A2 medium containing different ...
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The objective of this study was to improve induction of embryogenesis in bread wheat microspores. In this study, some F1 hybrids i.e M85-6 × 90, M85-8 × 90, mv17 × shiroudi, mv17 × kavir and kavir × bam were used. Microspores were cultured in A2 medium containing different amounts of maltose (60¬ & ¬90 ¬g/lit) depending on the genotype used. Analysis of variance for embryogenesis and regenerable embryos showed highly significant difference between hybrids but there was no significant difference between media (A2-60¬ &¬ A2-90) and interaction effects. M85-6 × 90, mv17 × Shiroudi and mv17 × kavir produced the highest ratio of embryogenesis among the hybrids. In regeneration phase, mv17 × Shiroudi and M85-6 × 90 had the highest frequency of regenerable embryos. The effect of 2,4-D as a novel stress for induction of microspores embryogenesis in Falat -known as the most responsive wheat cultivar to microspore culture technology, was investigated. Microspores were subjected to 2,4-D at 3 concentrations including 15, 25, 35 mg/l for 30 minutes while microspores without any stress treatment were used as the control. The embryogenesis of microspores stressed with 2,4-D were better than control. The highest yield of embryogenesis was produced at 15 mg/l 2,4-D. The most regenerated embryos were obtained in 15 & 25 mg/l of 2,4-D. According to the results obtained, 2,4-D is introduced as a new stress for induction of embryogenesis in microspores of wheat.
