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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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Results in the determination of some Hosta varieties by the method of isoelectric focusing
90-92.Views:111Mass propagation of 5 newly introduced Hosta varieties was carried out by the method of micropropagation. Our aim was to determine exact variety specificity after the micropropagation period in the pattern of peroxidase isoenzymes by isoelectric focusing in pH 3-9 range and to determine that phenological phase of mother plant in which the isoenzyme pattern of mother plant can safely be comparable to the isoenzyme pattern of micropropagated descendants. The isoenzyme patterns of descendants were similar to the mother plants of the same hybrid lines. The older leaves seemed to be not so suitable for examination than newly developed ones despite of the higher activity of peroxidase enzymes. There were big differences in isoenzyme patterns of leaves in different phenological phases. With this quick and easy method Hosta varieties could be selected already in the very early stage of micropropagation.