| Juhua is the flower of Chrysanthemum morifolium Ramat., a traditional Chinese herbal medicine plant and it occupies an important position in the Chinese medicine enterprises. Nowadays, the cultivation methods of C. morifolium are gradually changing and improving with the development of GAP and the people's awareness of the relationship between traditional medicine and health, therefore, the cultivation trends to much more scientific. Macroelements are integrant for the growth and metabolism of C. morifolium, its rational application can improve not only yield but also characteristic of the plant. This study is to research the effects of macroelements deficiency on the growth, primary and secondary metabolism, and investigate the secondary metabolites with HPLC and FTIR approaches in the different organs and diverse growth stages of C. morifolium. The purpose of this issue is to provide theoretical basis for the GAP and the secondary metabolism research. Main results are as follow.1. N deficiency in different life stages could decrease the yield of C. morifolium, and more decrease of the N application, more reduce of the yield. N deficiency in whole life stage could increase the flavonoid and soluble sugar contents both in leaves and flowers. Moreover, N deficiency could affect the solubility of the flavonoid. The content of middle-microelements significantly related to the flavonoid, for instance, among them Ca related positively to the flavonoids in three different treatments.2. N deficiency could influence the kinds and contents of secondary metabolites in HPLC figure print of leaves, and that related to the life stage of C. morifolium. And the results of FTIR show that the effect of N deficiency in different life stages was at the wavenumber of 1516 and 1325 cm-1, which was the characteristic area of flavonoids.3. N deficiency in whole life stage affected the growth of plant, and the effect was also found in the development process of the lateral branch. N deficiency improved the transpiration rate, stomatal conductance and intercellylar CO2 concentration, but decreased the potosythetic rate in the same time. In addition, N deficiency can accelerate the development of new roots, but that still lower than CK, the growth of root in N2 (0.2 g N/kg soil) was growth of the worst in three treatments, which suggest that the root could develop extremely in the reproductive stage.4. N deficiency could not affect the content of cinnamic acid both in leaves and flowers, but it could improve the content of P-coumaric acid in flower. N1(0.05 g N/kg soil) could improve the phenylalanine in leaves, while N2 (0.2 g N/kg soil) could improve it in flowers. The results of PCA show that the flavonoid synthesis process was different with CK compared to N1 (0.05 g N/kg soil) and N2 (0.2 g N/kg soil) treatments. The main flavonoid synthesis process was documented by PAL under normal condition, however, the dominance was changed to PAL and 4CL (C4H and 4CL) under N2 (N1) condition.5. P deficiency in different life stages could decrease the yield of C. morifolium, and more decrease of the P application, more reduce of the yield. P1 (0.05 g P2O5/kg soil) could increase the content of flavonoid and soluble sugar in leaves and flowers. P deficiency lead to the improvement of flavonoids in flowers, and then the effect appeared in the leaves in reproductive stage. The content of nutrients significantly related to the flavonoid, for instance, Ca positively related to the flavonoids in three treatments.6. P deficiency could not affect the FTIR figure print of C. morifolium. The effect of P deficiency appeared in the reproductive stage in HPLC figure print, and P deficiency in different life stages increased the content of flavonoids.7. P deficiency decreased the growth of lateral branch and the development of buds in these branches, which reduced to the decreasing of the yield. P deficiency could not affect the content of chlorophyll, but increase the transpiration rate, stomatal conductance and of leaves in vegetable life stage. In reproductive stage, P deficiency decreased the transpiration rate, stomatal conductance and intercellylar CO2 concentration, but increased the potosythetic rate in the same time. P1 (0.05 g P2O5/kg soil) accelerated the development of root, which opposite to P2 (0.1 g P2O5/kg soil). P deficiency could increase the content of flavonoids in root of C. morifolium.8. P deficiency could influence the secondary metabolites in phenolic synthesis pathway in plant of C. morifolium, and the regression equation between flavonoid and phenylalanine content was Y=-294.46X+150.66 (R2=0.9205, P<0.01) in CK and Y =42.62X+2.49 (R2=0.9564, P<0.01) in P deficient treatment (Y, flavonoid content; X, phenylalanine content). There were two principal components to control the phenolic major synthesis process, that was principal 1 (phenylalanine, PAL, cinnamic acid and p-coumaric acid) (88.17%) and principal 2 (4CL, C4H) (9.64%) domination under normal growth of C. morifolium. However, under P deficiency condition, the principal components were principal 1 (phenylalanine, cinnamic acid and p-coumaric acid and C4H) (81.46%) and principal 2 (PAL)(18.53%)domination, respectively. The influence of P deficiency on phenolic major synthesis pathway related to the change of PAL and C4H activities. 9. K deficiency decreased the flavonoid content by 31.4% in flower of C. morifolium. The microelements related to the flavonoid significantly, but the regression equation was different with each other. K deficiency decreased the protein content in leaves, but did not affect the protein in flowers. K1 (0.05 g K2O/soil) increased the soluble sugar in leaves obviously by 55.9%,22.1% and 21.1%.10. The FTIR figure print of flowers and leaves were same in different K deficience condition, which suggested that the K deficiency did not affect the function group both in flower and leaves. The HPLC figure print was different in leaves in different life stages, but the difference was not significantly. The HPLC figure print of leaf was significantly different with flower.11. K deficiency decreased the growth of lateral branch and the development of buds in these branches, and then decreased the yield of the plant. K deficiency increased the transpiration rate and stomatal conductance in leaves of vegetative life stage, however, in reproductive stage, K deficiency decreased the transpiration rate, stomatal conductance and intercellylar CO2 concentration, but increased the potosythetic rate. K deficiency decreased the development of the root.12. K deficiency decreased the flavonoid and chlorogenic acid contents slightly in flower of C. morifolium. The regression equations between phenylalanine, cinnamic acid, p-coumaric acid and flavonoid content were Y=285.4118X2-286.1689X+79.8951(R2 =0.9375, P<0.05), Y=2.4368X+3.0380 (R2=0.9809, P<0.001) and Y=0.2476X+ 5.5720 (R2=0.9929, P<0.0001) under K deficiency (Y, flavonoid content; X, phenylalanine, cinnamic acid and P-coumaric acid, respectively). Principal component analysis illustrate that K deficiency did not influence the preliminary flavonoid synthesis pathway. There were two major principals from chorismate to coumaryl Co A synsthesis process and the substrate was the domination principal in both K deficient (88.36% and 10.57%) and sufficient treatments (88.71% and 9.64%).
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