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Influence of antiobiotics on NAA- induced somatic embryogenesis in eggplant (Solanum melongena L. cv. Embil)
88-95.Views:156The influence of increasing concentrations of naphthaleneacetic acid and the antibiotics cefotaxime, timentin, kanamycin, and hygromycin on eggplant (Solantun melongena L. cv. Embil) somatic embryogenesis was investigated. Cotyledon explants were excised from 16 to 20 days old in vitro grown seedlings. NAA promoted somatic embryogenesis, although its concentrations had no influence on the mean number of embryos. Callusing decreaSed significantly with increasing NAA concentrations. Morphogenesis was stopped with 50 to 100 mg L-1 kanamycin and 7.5 to 15 mg L-1 hygromycin. Although early globular embryos were observed up to 15 mg L-1, further embryo development was inhibited at 10 mg L-1. Interestingly, cefotaxime (250 and 500 mg L-1) promoted a marked effect on enhancing fresh weight of calli, accompanied by decrease in embryo regeneration, whereas timentin concentrations (150 and 300 mg L-1) did not affect embryo differentiation as compared to the control treatment.
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Ultrastructural and biochemical aspects of normal and hyperhydric eucalypt
61-69.Views:211Hyperhydricity was observed throughout in vitro multiplication phase of a Eucalyptus grandis clone. Ultrastructural approach of tissue and cell differentiation, izoenzyme patterns, binding protein (BiP) expression, and pigment content were performed. Hyperhydric tissues showed a reduction in cell wall deposition, reduction of membranous organelles, higher cell vacuolation, and more intercellular spaces than its normal counterpart. Additionally, several vesicles were present in hyperhydric cells suggesting the occurrence of organelle autophagy by autophagic vacuole. Lower pigment content, intercellular spaces on the epidermis and the induction of a molecular chaperone (BiP) were observed in hyperhydric phenotype. Evidences of schizolysigenous process of intercellular space formation are compatible with a stress condition. Although plastoglobulli were observed in normal and hyperhydric chloroplasts, they were more evident in the normal ones. Abnormal stomata also reflected a disruptive situation and morphogenesis disturbances which would difficult plant acclimatization. Further observation of the epidermis ultrastructure allows us to conclude that the presence of intercellular spaces on its surface may be constraining the recovery and development of hyperhydric plants. Similarly to BiP, other proteins such as esterase (EST), acid phosphatase (ACP), malate dehydrogenase (MDH) and peroxidase (PDX) are possible to be used as stress markers in in vitro conditions. Our results confirm earlier findings about negative effects of hyperhydricity on in vitro plant morphogenesis and ultrastructure, which in eucalypt is associated with a stressful condition contributing to lower propagation ratios.
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Dr. Ottó Orsós, the forgotten Hungarian pioneer in plant tissue culture
9-13.Views:161The knowledge of tissue culture deserves attention in respect of understanding the development of universal biology. This study intends to contribute to the past of the plant tissue culture by such data of the history of science which have been unprocessed so far. It seems that the life-work of the Hungarian biologist, Dr. Ottó Orsós is a missing and essential link between those early plant hormone researchers and the representatives of the pioneers of tissue culture schools who have contributed substantially to the development of the modern in vitro plant morphogenesis and plant cell biology. Orsós cultured kohlrabi tuber cubes on White culture medium in a sterile manner. This way, he could efficiently direct the in vitro morphogenesis of the kohlrabi, the regeneration of its shoot and root, and the formation and steps to subculture of pure callus tissues in 1938. He supported the correctness of its statements by means of detailed anatomical examinations. Orsós successfully rooted and aclimatized complete regenerated plants. We may as well call the above system — in remembrance of the creators of the original concept — "Haberlandt-Orsós model". Between the publishing of his main paper in 1938 and 2003, a period of 65 years has lapsed. On the occasion of this anniversary, we bow before this forgotten pioneer.
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In vitro plant regeneration from immature embryo axis and cotyledons of common bean (Phaseolus vulgaris L.)
93-97.Views:127Phaseolus vulgaris L. is the most important economic species within the genus Phaseolus. It is grown in all parts of the world. Genetic improvement by conventional breeding has met considerable success, although production of hybrids between species within the genus has been limited due to sexual incompatibility. Recent advances in tissue culture have offered the opportunity to produce cultivars, which could not be obtained by conventional breeding methods. The use of tissue culture and genetic engineering is viewed as a logical approach to improve bean production. Gene transfer techniques will have a great impact on legumes. Although the concept of cell totipotency is widely proved, in vitro morphogenesis has not yet been achieved for a large number of cultivated beans. Regeneration protocols are strongly influenced by the genotype. In tissue and cell culture of beans, the factors controlling shoot morphogenesis and somatic embryogenesis are still unknown. The reported data suggest a possible way for future research.
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Influence of different growth regulators on the in vitro morphogenesis of an ornamental variety of carnation
55-57.Views:137Callus formation, as a prerequisite for the induction of somaclonal variability, was achieved successfully with certain molar ratios between 2,4-dichlorophenoxyacetic acid and benzyladenine. Regeneration of new plants from shoot apex meristems could be significantly improved by the combined addition of very low amounts of indolebutiric acid, benzyladenine and gibberelic acid, dissolved in the Murashige-Skoog nutrient medium. These in vitro treatments may contribute to a more efficient micropropagation of the Rimini variety of carnation.
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Organogenesis in eggplant (Solanum melongena L. cv. Embu) as affected by antibiotics and growth regulators
76-82.Views:214The influence of antibiotics (cefotaxime, timentin, kanamycin and hygromycin) and growth regulators (indolacetic acid and 6-benzylaminopurine) was evaluated on eggplant organogenesis. Solanum melongena hypocotyl segments (6 to 10 mm length), taken from 16 to 20-days in vitro grown seedlings, were used as explants. The basic medium was composed by MS salts, Gamborg vitamins and 2% sucrose, solidified with agar 0.8% and pH adjusted to 5.7±0.2. Morphogenesis was impaired at 50 to 100 mg L-I kanamycin and 7.5 mg L-1 hygromycin. Both Timentin and cefotaxime reduced the frequency of regenerating explants meanwhile hyperhydricity was not affected. A decrease in root regeneration was observed with increasing cefotaxime concentrations, although, timentin had no effect on root regeneration, as compared to the control treatment. Interestingly, the number of adventitious roots was more noticeable at 0.25 mg L-I IAA plus 0.5 mg L-1 BAP. However, if just IAA was added led to higher number of regenerated roots compared to other treatments.