Asad Masoumiasl; Ameneh Aryiaeineghad; Masud Dehdeari
Abstract
Introduction: German (Matricaria chamomilla L.) and Shirazi (Matricaria recutita L.) chamomiles are the most important medicinal plants of the Astraceae family which are used in the pharmaceutical, health, food and cosmetics industries. Production of this plant has been undertaken in Iran mainly in Isfahan, ...
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Introduction: German (Matricaria chamomilla L.) and Shirazi (Matricaria recutita L.) chamomiles are the most important medicinal plants of the Astraceae family which are used in the pharmaceutical, health, food and cosmetics industries. Production of this plant has been undertaken in Iran mainly in Isfahan, Kohgiluyeh and Boyerahmad, Golestan and Hamedan provinces. In vitro propagation of plants have higher potential to produce qualified natural products, restoring and preserving of endangered plants, induction of somaclonal variation, industrial reproduction, valuable secondary metabolites and increased active ingredients. Researchers reported successful micropropagation system for five chamomile varieties on MS medium contained 0.01 mg/l NAA and 2.5 mg/l kinetin. The aim of this study was to investigate the effects of stem (with and without node), leaf and cotyledon explants and different plant growth regulators on direct regeneration of German and Shirazi chamomiles. Although the Shirazi chamomile is native to Iran, but German chamomile is Iran non-indigenous cultivar. By our knowledge, there have been no comparison reports about responses of these cultivars to tissue culture.
Materials and Methods: This research was performed in the central laboratory of Agriculture Faculty in Yasouj University. Seeds were provided from Pakan-Bazr institute, Isfahan. Chamomile seeds were disinfected by ethanol (70%) for 5-10 min and sodium hypocholorite 3% for 5-12 min and then washed for several times by distilled water. Then, seeds were sown on MS medium for germination. After 2-3 weeks, seedlings were grown and then planted in MS medium supplemented with hormonal combinations of NAA at two levels (0.1 and 0.5 mg/l), kinetin at three levels (2, 2.5 and 3 mg/l) and Zeatin, BAP and 2ip at three levels (0.5, 1 and 1.5 mg/l). The experiment was performed in a completely randomized design with four replications. Factors included explants, cultivars and hormonal combinations. In this experiment, traits such as stem induction percentage, stem length, stem fresh and dry weight root induction percent, root length and stem fresh and dry weight were measured. Statistical analysis was performed using SAS software (version 9.1). In order to test the normality and perform mean comparisons, Minitab 14 and MSTAT-C software was used and excel software was used for drawing diagrams.
Results and Discussion: For all traits except stem induction percent and root dry weight, triple interaction of explants, hormonal combinations and cultivar were significant at 1% level. Triple mean comparisons for cultivar, explant and hormonal combinations shows that the highest mean for stem height, stem fresh weight, stem dry weight, root induction percent, root height and root fresh weight was obtained in Shirazi Chamomile cultivar with cotyledon explants in hormonal combinations of 0.1 mg/l NAA and 1.5 mg/l 2ip. The results showed that the best explants in both chamomiles for direct regeneration were stem (whit node) and cotyledon with 78.75% and 75% regenerations, respectively. Also the best genotype and hormone combination were Shirazi chamomile and MS medium supplemented with 1.5mg/l 2ip with 0.1mg/l NAA. To determine the best hormonal combination for root regeneration from direct regeneration, the stems were embedded in medium contained different concentrations of IBA. The first signs of rooting production were observed after 5-7 days. After completing the roots formation (4 weeks after transferring), the rooting percentage, root length, fresh and dry weight of roots were measured. Based upon variance analysis, effect of triple interactions of IBA, explants and cultivar on all traits were not significant, but the effect of IBA for all traits was significant at 1% level. According to the results of mean comparisons for effects of IBA on root traits in direct regeneration of chamomile, the highest percentage of root regeneration (73.75 %), the highest root length (6.60 cm), root fresh weight (174.167 mg) and the highest root dry weight (16.425 mg) were obtained from medium contained 0.5 mg/l IBA. Root differentiation was influenced by auxin (0.5 mg/l IBA) or spontaneously. Regenerated plantlets were transferred to pots contained sterilized soil (3:1:1 mixture of soil: sand: leaf compost). For plant adaptation to natural conditions, glassy caps were used. After adaptation, the caps were removed and the plants were transferred into a growth chamber. Previous studies reported that MS medium supplemented with 0.2 to 1 mg/l of BA and 2 mg/l of NAA induced adventitious bud formation and shoot development in leaf explants of Roman Chamomile. A higher number of adventitious buds were observed at the proximal end of the explants. Plantlets were rooted on MS medium supplemented with 0.1 mg/l of IBA and successfully weaned in vivo.
Conclusion: Based on the results of this research, chamomile showed relevant response to direct regeneration.
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.
Mohammad Gerdakaneh
Abstract
The present investigation was conducted to study the effects of type and concentration of auxin and cytokinin on callus induction of three strawberry (Kurdistan, Parose and Camarosa) cultivars. For this purpose, leaf blade, nodal, petiole, receptacle, stamen and flower bud explants were cultured on Murashige ...
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The present investigation was conducted to study the effects of type and concentration of auxin and cytokinin on callus induction of three strawberry (Kurdistan, Parose and Camarosa) cultivars. For this purpose, leaf blade, nodal, petiole, receptacle, stamen and flower bud explants were cultured on Murashige and Skoog (MS) medium at three separate experiments included 2, 4 dichlorophenoxyacetic acid (2,4-D) at 2, 3, 4, 5 and 6 mg/L, naphthalene acetic acid (NAA) at 2, 3, 4, 5 and 6 mg/L and different concentrations (0, 0.5, 1.0, and 2.0 mg/L) of benzyl adenine (BA) combination with 4 mg/L NAA. The concentration and type of growth regulators, cultivar and explant type were found critical to the callus induction and callus physical appearance. All explants exception of receptacle incubated on medium formed callus. The leaf and nodal were the most responsive explant to produce callus. MS medium supplemented with combination of auxin and cytokinin were evidently higher than those in the single auxin treatments. 4.0 mg/L NAA + 1 mg/L BA yielded the highest percentage of callus in all types of explants. In this medium, the highest callus induction (98.50%) was recorded in nodal explants of Kurdistan cultivar.
Mohammad Gerdakaneh; Ali Akbar Mozafari; Abolmohsen Hajiamiri
Abstract
Present study was conducted to consider the effects of different concentrations of 2,4-D and BA on somatic embryogenesis induction, development and maturation of three strawberry cultivars (Kurdistan, Parose and Camarosa). For this purpose, leaf blade, nodal, petiole, stamen and flower bud calli were ...
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Present study was conducted to consider the effects of different concentrations of 2,4-D and BA on somatic embryogenesis induction, development and maturation of three strawberry cultivars (Kurdistan, Parose and Camarosa). For this purpose, leaf blade, nodal, petiole, stamen and flower bud calli were cultured on Murashige and Skoog (MS) medium supplemented with 2, 4 dichlorophenoxyacetic acid (2,4-D at 0.25, 0.5, 1 and 2 mg/l ) as well as the combination with different concentrations (0.0, 0.25, 0.5, and 1 mg/l) of benzyl adenine (BA). The concentration and kind of growth regulators, cultivar and explant had great importants on strawberry somatic embryogenesis induction, development and maturation. Results obtained from the studies revealed that all explants with the exception of petiole and stamen incubated on medium formed embryonic calli. MS medium supplemented with combination of auxin and cytokinins were evidently higher than those in the single auxin treatments. 1 mg/l 2,4-D + 0.25 mg/l BA yielded the highest percentage of embryonic calli and number of globular embryo stage and 0.5 mg/l 2,4-D + 0.25 mg/l BA yielded the highest number of embryo cotyledonary in all types explants. The leaf explant calli and Parose cultivar were the most responsive to produce to somatic embryogenesis induction, development and maturation.