The gene encoded for tryptophan decarboxylase (TDC), which is the key enzyme in terpenoil indole alkaloids pathway, was targeted to different subcellular compartments and stably expressed in transgenic tobacco (Nicotiana tabacum L.) plants at the levels detected by Western blot and tryptamine accumulation analysis. It was shown that the TDC was located in subcellular compartments, the chloroplasts and cytosol. The recombinant TDC targeted to chloroplasts and cytosol in tobacco plants was effectively expressed as soluble protein by Western blot analysis and enzymatic assay. The level of tryptamine accumulation in chloroplast was higher than that in cytosol and very low in vacuole and endoplasmic reticulum (ER) to be hardly detected by Western blot analysis. It was indicated that the highest amount of tryptamine was in chloroplasts, lower in endoplasmic reticula and the lowest in vacuoles as compared to those in wild type plants. The TDC targeted to different subcellular compartments of tobacco plants and its expression level were studied by different nucleotide sequences coding signal peptides at 5'-end of tdc gene in order to know the effects of the TDC in compartmentation on its functionality.
Potted seadlings of Pinus koraiensis , Fraxinus mandshurica,Juglans mandshurica,Tilia amurensis, and Quercus mongolica ,which are five dominant species in the Korean pine broadleaf forest at Changbai mountain,were grown in different soil moistures.We designed three soil moisture scenarios:85%~100%(high water,CK),65%~85% (medium water,MW) and 45%~65% (low water,LW) of field water holding capacity.The results show that characteristics of typical drought resistance on the leaves are significantly developed.The net photosynthetic rate and water use efficiency of F. mandshurica were higher compared with CK at MW.The net photosynthetic rate and water use efficiency of other 4 tree species at CK were lower than those at MW and LW.The transpiration rate of 5 tree species responses differently to various soil water status.
Strictosidine synthase (STR) is a key enzyme involved in the biosynthesis of terpenoid indole alkaloids (TIA) by condensing tryptamine and secologanin into strictosidine. The transgenic tobacco plants targeting STR to subcellular compartments were established to express STR in chloroplast, vacuole and endoplasmic reticulum (ER) by the tobacco stable transformation. It was shown that STR was effectively expressed in the above subcellular compartments by Western blot analysis and STR enzymatic assay. In vitro , STR enzymatic assay was measured indirectly by fluorimetrically detecting depletion of tryptamine feeding on secologanin in the reaction mixture. The tryptamine were completely depleted by STR in the crude extract of leaves of transgenic tobacco plants targeting and expressing STR in the chloroplast, vacuole and ER, which ascertained the STR functionally targeted to the three subcellular compartments. To confirm STR correct targeting and expressing in chloroplast, the chloroplasts were isolated and the fractions of purified chloroplasts were analyzed by Western blot. The hypothesis of STR correct targeting to the chloroplast was tested. The results have implications on our understanding of the complex intracellular trafficking in metabolic intermediates of TIA biosynthesis.
Eco-physiological responses of seedlings of eight species, Pinus koraiensis, Picea koraiensis, Larix olgensis, Populus ussuriensis, Betula platyphylla, Tilia amurensis, Traxinus mandshurica and Acer mono from broadleaved/Korean pine forest, to elevated CO2 were studied by using open-top chambers under natural sunlight in Changbai Mountain, China in two growing seasons (1998-1999). Two concentrations of CO2 were designed: elevated CO2 (700 祄olmol-1) and ambient CO2 (400 祄olmol-1). The study results showed that the height growth of the tree seedlings grown at elevated CO2 increased by about 10%-40% compared to those grown at ambient CO2. And the water using efficiency of seedlings also followed the same tendency. However, the responses of seedlings in transpiration and chlorophyll content to elevated CO2 varied with tree species. The broad-leaf tree species were more sensitive to the elevated CO2 than conifer tree species. All seedlings showed a photosynthetic acclimation to long-term elevated CO2.