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.
To understand influence of litters on the emission/absorption of CO2, N2O and CH4 in broadleaved/Korean pine forest in Changbai Mountain, fluxes of soil CO2, N2O and CH4 were measured by closed static chamber technique, from Sept 3, 2002 to Oct 30, 2003 in two types of soil ecosystems, of which one was covered with litters on the surface soil, and the other had no litters. The results showed that litters had significant influences on CO2, N2O and CH4 fluxes (p<0.05). Their diurnal change patterns of plot with litters and litter-free plot were similar, and they all showed emission/absorption peak at 18:00. The diurnal change fluxes of CO2 and N2O of plot with litters were significantly higher than those of the litter-free plot, while the diurnal flux of CH4 of plot with litters was lower than that of litter-free plot. The fluxes of CO2, N2O, and CH4 showed the similar seasonal patternsfor both plots. The fluxes of CO2, CH4 showed their peak fluxes in June, but the fluxes of N2O showed its peak emissions in August. The annual fluxes of CO2 and N2O of plot with litters were significantly higher than those of the litter-free plot, while the annual flux of CH4 of plot with litters was lower than that of litter-free plot. Keywords Flux - CO2, N2O and CH4 - Seasonal variation - Diurnal variation CLC number S718.5 Document code A Foundation item: The study was supported by innovation research project of Institute of Appiied Ecology, Chinese Academy of Sciences (SCXZD0101-02) and National Natural Science Foundation of China (30271068)Bigraphy: XIAO Dong-mei (1979-), female, master of Institute of Applied Ecology. Chinese Academy of Sciences, Shenyang 110016. P. R. China.Responsible editor: Song Funan
