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The influence of kinetin and sucrose on (60%), Hill activity (about 3-fold), and "C-fixation from NaH"CO3 (about 20%). On the other hand, the presence of sucrose in the medium reduced the chlorophyll content by about 30% and "C-fixation from NaH"4CO3 in the soluble fraction by about 60%. A possible correlation between the influence of kinetin on sugar uptake and the effect of kinetin on "C-fixation from NaH"CO3 was discussed.
During the first 8-12 days of cultivation in a nutrient solution containing IAA, inositol and kinetin freshly isolated carrot root explants develop into a chlorophyllous and photosynthetically active tissue culture. Electron microscopy, low temperature pigment absorption spectra and fluorescence induction profile recording as well as the determination of the activity of some enzymes (ribulosebisphosphate carboxylase, phosphoenolpyruvate carboxylase) and CO2-fixation experiments were carried out. Based on the results, a sequence of developmental stages of the photosynthetic system will be proposed. During the autotropic period from about the 20th day of culture onward the light reaction system of these tissue cultures is quite comparable to that of carrot leaves; however, some differences in the CO2-fixation mechanism were observed.
Investigations were performed on growth phase-dependent EcoRII site-specific DNA methylation of the carrot genome during primary culture to elucidate physiological aspects of genome DNA variability in tissue culture. While DNA methylation of the root cambium and the secondary phloem and petioles of carrot leaves were strikingly different, the methylation level of the secondary phloem seemed to be independent of cultivar origin, the age of the plants and the extent of secondary root growth. As was shown earlier a change in the differentiated state of the secondary phloem by tissue culture leads to changes in genome modification. Whereas de novo methylation was observed during the first 2 weeks of growth initiation, the results presented demonstrate genome de-methylation during the transition to stationary growth indicating differential εnome methylation during different phases of culture. The presence of kinetin in the nutrient medium of the primary culture was found to be antagonistic to changes in genome modification in general. De novo methylation and subsequent de-methylation of the carrot genome are discussed as gross changes obviously essential to molecular genome differentiation during tissue culture.
Chromoplasts, which exist in the ceils of freshly isolated carrot root explants, seemed to be transformed in thylakoid containing plastids, and chlorophyll formation was initiated if the explants were cultured in a liquid medium containing inositol and IAA as a hormonal supplement. This process was intensified when kinetin was also added, but no dependence on a sucrose supply could be found.A sucrose supply of 2% in conjunction with the combination of all three hormones, however, was needed to achieve maximal thylakoid formation including stacking in individual chloroplasts and for the very extensive chloroplast multiplication in explants growing with high cell division activity. It should be noted that the number of plastids per cell is strongly increased by the sucrose supplement which leads also to starch accumulation. However, no transformation into chloroplasts occurred without the hormonal stimulus.
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