We describe an instrument that allows the rapid measurement of fluorescence lifetime-resolved images of leaves as well as sub-cellular structures of intact plants or single cells of algae. Lifetime and intensity fluorescence images can be acquired and displayed in real time (up to 55 lifetime-resolved images per s). Our imaging technique therefore allows rapid measurements that are necessary to determine the fluorescence lifetimes at the maximum (P level) fluorescence following initial illumination during the chlorophyll (Chl) a fluorescence transient (induction) in photosynthetic organisms. We demonstrate the application of this new instrument and methodology to measurements of: (1) Arabidopsis thaliana leaves showing the effect of dehydration on the fluorescence lifetime images; (2) Zea mays leaves showing differences in the fluorescence lifetimes due to differences in the bundle sheath cells (having a higher amount of low yield photosystem 1) and the mesophyll cells (having a higher amount of high yield photosystem 2); and (3) single cells of wild type Chlamydomonas reinhardtii and its non-photochemical quenching mutant NPQ2 (where the conversion of zeaxanthin to violaxanthin is blocked), with NPQ2 showing lowered lifetime of Chl a fluorescence. In addition to the lifetime differences referred to in (1) and (2), structural dependent heterogeneities in the fluorescence lifetimes were generally observed when imaging mesophyll cells in leaves. and O. Holub ... [et al.].
At chilling stress, the contents of photosynthetic pigments decreased significantly in maize, but in wheat the contents of chlorophyll (Chl) remained unchanged whereas the contents of total carotenoids (Car) increased. In both species the contents of α+β carotene and lutein + lutein-5,6-epoxide remained unaffected, but the de-epoxidation state involving the components of the xanthophyll cycle increased. Under chilling stress the photosynthetic electron transport also displayed a general failure in maize but in wheat only photosystem (PS) 2 coupled to the water oxidation complex was inhibited. Moreover, in stressed maize the quinone pool decreased, while the low and high potential forms of cytochrome b559 increased. In wheat only the contents of cytochrome b559LP decreased. Peroxidation of acyl lipids in the chloroplast lamellae became more distinct in chilling stressed maize but could also be detected in wheat. Thus in chilling stressed maize prevails an impairment of the acceptor site of PS2 while in wheat photodamage is restricted to the electron donation pathway from water to P680 or to the oxygen evolving complex. and F. C. Lidon ... [et al.].
The influence of chilling (8 °C, 5 d) at two photon flux densities [PFD, L = 200 and H = 500 μmol(photon) m-2 s-1] on the gas exchange and chlorophyll fluorescence was investigated in chilling-tolerant and chilling-sensitive maize hybrids (Zea mays L., K383×K130, K185×K217) and one cultivar of field bean (Vicia faba L. minor, cv. Nadwiślański). The net photosynthetic rate (PN) for the both studied plant species was inhibited at 8 °C. PN of both maize hybrids additionally decreased during chilling. Changes in the quantum efficiency of PS2 electron transport (ΦPS2) as a response to chilling and PFD were similar to PN. Measurements of ΦPS2/ΦCO2 ratio showed that in field bean seedlings strong alternative photochemical sinks of energy did not appear during chilling. However, the high increment in ΦPS2/ΦCO2 for maize hybrids can indicate reactions associated with chill damage generation. At 8 °C the non-photochemical quenching (NPQ) increased in all plants with chilling duration and PFD. The appearance of protective (qI,p) and damage (qI,d) components of qI and a decrease in qE (energy dependent quenching) took place. NPQ components of field bean and maize hybrids differed from each other. The amount of protective NPQ (qE + qI,p) components as part of total NPQ was higher in field bean than in maize hybrids at both PFD. On 5th day of chilling, the sum of qE and qI,p was 26.7 % of NPQ in tolerant maize hybrids and 17.6 % of NPQ in the sensitive one (averages for both PFD). The increased PFD inhibited the ability of all plants to perform protective dissipation of absorbed energy. The understanding of the genotypic variation of NPQ components in maize may have implications for the future selection of plants with a high chilling tolerance. and J. Kościelniak, J. Biesaga-Kościelniak.
High level of phosphoenolpyruvate carboxylase (PEPC) gene was stably inherited and transferred from the male parent, PEPC transgenic rice, into a female parent, japonica rice cv. 9516. Relative to the female parent, the produced JAAS45 pollen lines exhibited high PEPC activity (17-fold increase) and also higher photosynthetic rates (about 36 %-fold increase). The JAAS45 pollen lines were more tolerant to photoinhibition and to photo-oxidative stress. Furthermore, JAAS45 pollen lines, as well as their male parent, were tested to exhibit a limiting C4 cycle by feeding with exogenous C4 primary products such as oxaloacetate (OAA). Thus the PEPC gene and photosynthetic characteristics of PEPC transgenic rice could be stably transferred to the hybrid progenies, which might open a new breeding approach to the integration of conventional hybridization and biological technology. and L. Ling, B. J. Zhang, D. M. Jiao.
Pot trial in greenhouse was conducted using cumulic cinnamon soil from North China to study the effects of zinc deficiency on CO2 exchange, chlorophyll fluorescence, the intensity of lipid peroxidation, and the activity of superoxide dismutase (SOD) in leaves of maize seedlings. Zn deficiency resulted in a reduction of net photosynthetic rate and stomatal conductance to H2O. The maximum quantum efficiency of photosystem 2 (PS2) and the PS2 activity were depressed, while the pool size of the plastoquinone molecules was not affected by Zn deficiency. The content of super oxygen anion radical (O2.-) and the intensity of lipid peroxidation as assessed by malonyldialdehyde content in Zn-deficient leaves were higher than those in Zn-sufficient leaves. The activity of SOD increased with Zn application. The adverse influence of Zn-deficiency on the light stage of photosynthesis is probably one of possible reasons for the limitation of photosynthetic capacity in maize leaves. and H. Wang, J. Y. Jin.
Etiolated leaves of three different species, maize, wheat, and pea, as well as a pea mutant (lip1) were used to compare the excitation spectra of protochlorophyllide (Pchlide) in the red region. The species used have different composition of short-wavelength and long-wavelength Pchlide forms. The relation between different forms was furthermore changed through incubating the leaves in 5-aminolevulinic acid (ALA), which caused an accumulation of short-wavelength Pchlide forms, as shown by changes in absorption and fluorescence spectra. This is the first time a comprehensive comparison is made between excitation spectra from different species covering an emission wavelength range of 675-750 nm using fluorescence equipment with electronic compensation for the variations in excitation irradiance. The different forms of Pchlide having excitations peaks at 628, 632, 637, 650, and 672 nm could be best measured at 675, 700, 710, 725, and 750 nm, respectively. Measuring emission at wavelengths between 675-710 nm gave an exaggeration of the short-wavelength forms and measuring at longer wavelengths gave for the pea leaves an exaggeration of the 672 nm peak. In general, an energy transfer from short-wavelength Pchlide forms to long-wavelength Pchlide forms occurred, but such an energy transfer sometimes seemed to be limited as a result of a discrete location of the Pchlide spectral forms. The excitation spectra resembling the absorption spectrum most were measured at an emission wavelength of 740 nm. Measuring the excitation at 710 nm gave higher intensity of the spectra but the short-wavelength forms were accentuated. and M. R. Amirjani, C. Sundqvist.
Incubation of maize NADP-malic enzyme with tetranitromethane (TNM) resulted in a total loss of enzyme activity. The loss of enzyme activity was not observed at pH 6.3 but at pH 8.0. NADP-malic enzyme was inactivated to almost 90 % by incubation with an 80-fold molar excess of TNM for 5 min at 30 °C. The substrate malate or Mg2+ alone gave no protection, while NADP provided considerable protection. NADP in the presence of malate and Mg2+ totally protected the enzyme activity, suggesting that tyrosine residue may be located at or near the active site of maize NADP-malic enzyme. The spectral analysis of the modified enzyme indicated that modification of at least one tyrosine residue per subunit resulted in complete loss of the enzyme activity. The fluorescence study of unmodified and modified enzymes postulated that essential tyrosine residue at maize NADP-malic enzyme is possibly involved in malate binding. and S. R. Rao, B. G. Kamath, A. S. Bhagwat.