Chloroplasts were isolated from spinach cultured in lead chloride-present, Ce3+-administered, cerium chloride-administered lead chloride-present Hoagland’s media or that of Hoagland’s media. The experimental study demonstrated the effects of cerium (Ce) on distribution of light energy and photochemical activities of spinach chloroplast grown in lead (Pb)-present media. It was observed that Pb2+ significantly inhibited photosynthesis in spinach, including light absorption, energy transfer from LHCⅡ to photosystem Ⅱ, excitation energy distribution from photosystem I to photosystem Ⅱ, and transformation from light energy to electron energy and oxygen evolution of chloroplasts, and decreased spinach growth. However, Ce3+ treatment to Pb2+-present chloroplasts could obviously improve light absorption and excitation energy distribution in both photosystems and increase activity of photochemical reaction and oxygen evolution of chloroplasts. The results suggested that Ce3+ under Pb2+ stress could maintain the stability of chloroplast membrane, and improve photosynthesis of spinach chloroplast, thus promote spinach growth.
The complex of Rubisco and Rubisco activase from LaCl3-, CeCl3- treated spinach in vivo is induced. SDS-PAGE result shows that the purified proteins from LaCl3-, CeCl3- treated spinach have not only large and small subunits (55 kD, 14.4 kD) of Rubisco, but also two large subunits of 45 kD and 41 kD near the large subunits of Rubisco. Native-PAGE shows that the purified proteins from LaCl3-, CeCl3- treated spinach have not only a band of Rubisco (560 kD), but also a band of about 1100 kD, about twice distant from Rubisco, which might be a complex of Rubisco and Rubisco activase. The purified enzyme activities from LaCl3-, CeCl3-treated spinach are 1.8 and 2.8 times that of the control,the intensities of absorption and fluorescence are sig- nificantly higher than that of the purified Rubisco from the control, and the total sulfhydryl groups and available sulfhydryl groups are 36—39 –SH per mol enzyme, 14—25 –SH per mol enzyme more than those of the purified Rubisco from the control, respectively. The CD spectra show that the secondary structure of the purified enzyme from LaCl3-, CeCl3 -treated spinach is very dif- ferent from the control. The enzyme activities from LaCl3-, CeCl3- treated spinach in vivo are 1.5 and 1.9 times those of the control.
HONG Fashui1, LIU Chao1, ZHENG Lei1, WANG Xuefeng1, WU Kang1, SONG Weiping1, Lü Shipeng1, TAO Ye2 & ZHAO Guiwen3 1. College of Life Sciences, Suzhou Universty, Suzhou 215006, China
Linolenic acid has great effects on the structure and function of chloroplast. The function of Ce^3＋ on the improvement of chloroplast photoreduction activity and oxygen evolution damaged by linolenic acid in spinach by in vitro investigation was studied. Results showed that adding Ce^3＋ to the linolenic acid treated chloroplast could greatly decrease the reduction linolenic acid exerted on the whole chain electron transport rate and the photoreduction activity of photosystem Ⅱ （PSII） and photosystem Ⅰ （PSI） as well as the oxygen evolution rate of chloroplast. It indicated that Ce^3＋ had the ability to relieve the inhibition of the photochemical reaction of chloroplast caused by linolenic acid to some extent.
Chloroplasts were isolated from spinach treated with taCl3, CeCl3, and NdCl3. Because of owning 4f electron characteristics and alternation valence, Ce treatment presented the highest enhancement in light absorption, energy transfer from LHC Ⅱ to PS Ⅱ, excitation energy distribution from PS Ⅰ to PS Ⅱ, and fluorescence quantum yield around 680 nm. Compared with Ce treatment, Nd treatment resulted in relatively lower enhancement in these physiological indices, as Nd did not have alternation valence. La treatment presented the lowest enhancement, as La did not have either 4f electron or alternation valence. The increase in activities of whole chain electron transport, PS ⅡDCPIP photoreduction, and oxygen evolution of chloroplasts was of the following order： Ce〉Nd 〉La〉 control. However, the photoreduction activities of spinach PS I almost did not change with La, Ce, or Nd treatments. The results suggested that 4f electron characteristics and alternation valence of rare earths had a close relationship with photosynthesis improvement.
Due to 4f electron characteristics and alternation valence, cerium involved in an oxidation-reduction reaction in plant, closely relating to photosynthesis. Our studies proved that cerium could promote photosynthesis and greatly improve spinach growth. However, the mechanism of promoting energy transfer and conversion by cerium remains unclear. Here we reported that the effects of Ce^3＋ on energy transfer and oxygen evolution in photosystem Ⅱ （PS Ⅱ ） isolated from spinach, which was related to 4f electron characteristics and alternation valence in Ce molecule. The methods of absorption spectrum, fluorescence spectrum were used in the research. Results showed that Ce^3＋ treatment at low concentration could suitably change PS Ⅱ mieroenvironment, increase the absorbance of visible light, improve the energy transfer among amino acids within PS Ⅱ protein-pigment complex, and accelerate energy transport from tyrosine residue to chlorophyll a. In summary, the photochemical activity of PS Ⅱ （fluorescence quantum yield） and its oxygen evolving rate were enhanced by Ce^3＋.
The effects of Ce^3 ＋ on the chloroplast senescence of spinach under light were studied. The results show that when the chloroplasts are illuminated for 1, 5 and 10 min with 500 μmol·cm^-2· min^-1 light intensity, the oxygen evolution rate is rapidly increased. When the chloroplasts are treated for 20, 30 and 40 min with 500 μmol·cm^-2·min^-1 light intensity, the oxygen evolution rate is gradually decreased. While spinach is treated with 16μmol·L^-1 Ce^3＋ , the rate of oxygen evolution of chloroplasts in different illumination time （1,5, 10, 20, 30, 40 min） is higher than that of control, and when illumination time is over 10 min, the reduction of the oxygen evolution rate is lower than that of control. It suggests that Ce^3＋ treatment can protect chloroplasts from aging for long time illumination. The mechanism research results indicate that Ce^3＋ treatment can significantly decrease accumulation of active oxygen free radicals such as O2^- and H2O2, and reduce the level of malondialdehyde （MDA）, and maintain stability of membrane structure of chloroplast under light. It is shown that the redox took place between cerium and free radicals, which are eliminated in a large number, leading to protect the membrane fiom peroxidating.
