Ultrasonic communication in frogs.

Among vertebrates, only microchiropteran bats, cetaceans and some rodents are known to produce and detect ultrasounds (frequencies greater than 20 kHz) for the purpose of communication and/or echolocation, suggesting that this capacity might be restricted to mammals. Amphibians, reptiles and most birds generally have limited hearing capacity, with the ability to detect and produce sounds below approximately 12 kHz. Here we report evidence of ultrasonic communication in an amphibian, the concave-eared torrent frog (Amolops tormotus) from Huangshan Hot Springs, China. Males of A. tormotus produce diverse bird-like melodic calls with pronounced frequency modulations that often contain spectral energy in the ultrasonic range. To determine whether A. tormotus communicates using ultrasound to avoid masking by the wideband background noise of local fast-flowing streams, or whether the ultrasound is simply a by-product of the sound-production mechanism, we conducted acoustic playback experiments in the frogs' natural habitat. We found that the audible as well as the ultrasonic components of an A. tormotus call can evoke male vocal responses. Electrophysiological recordings from the auditory midbrain confirmed the ultrasonic hearing capacity of these frogs and that of a sympatric species facing similar environmental constraints. This extraordinary upward extension into the ultrasonic range of both the harmonic content of the advertisement calls and the frog's hearing sensitivity is likely to have co-evolved in response to the intense, predominantly low-frequency ambient noise from local streams. Because amphibians are a distinct evolutionary lineage from microchiropterans and cetaceans (which have evolved ultrasonic hearing to minimize congestion in the frequency bands used for sound communication and to increase hunting efficacy in darkness), ultrasonic perception in these animals represents a new example of independent evolution.

Folding studies of two hydrostatic pressure sensitive proteins.

High hydrostatic pressure combined with various spectroscopies is a powerful technique to study protein folding. An ideal model system for protein folding studies should have the following characteristics. (1) The protein should be sensitive to pressure, so that the protein can be unfolded under mild pressure. (2) The folding process of the protein should be easily modulated by several chemical or physical factors. (3) The folding process should be easily monitored by some spectroscopic parameters. Here, we summarized the pressure induced folding studies of two proteins isolated from spinach photosystem II, namely the 23-kDa and the 33-kDa protein. They have all the characteristics mention above and might be an ideal model protein system for pressure studies.

Manipulated photocurrent generation from pigment-exchanged photosynthetic proteins adsorbed to nanostructured WO3-TiO2 electrodes.

A novel photoelectrode (PE) consisting of the pigment-exchanged photosynthetic reaction center (RC) trapped on the mesoporous WO3-TiO2 film was fabricated to facilitate bio-photoelectric conversion by manipulating the excitation relaxation of the proteins.

The observation of ultrafast excited-state dynamical evolution in B800- partially or completely released LH2 of Rhodobacter sphaeroides 601 at room temperature.

Photodynamics of two kinds of peripheral antenna complexes (LH2 of Rhodobacter sphaeroides, native LH2 (RS601) and B800-released LH2 where B800-BChls were partially or completely removed with different pH treatments), were studied using femtosecond pump-probe technique at different laser wavelengths. The obtained results for these samples with different B800/B850 ratios demonstrated that under the excitation around B800 nm, the photoabsorption and photobleaching dynamics were caused by the direct excitation of upper excitonic levels of B850 and excited state of B800 pigments, respectively. Furthermore, the removal of B800 pigments had little effect on the energy transfer processes of B850 interband/intraband transfer.

Tyrosine residues of the extrinsic 23 kDa protein are important for its interaction with spinach PSII membranes.

The 1, 4, and 8 tyrosine (Tyr) residues on the PSII extrinsic 23 kDa protein were modified with 5, 10 or 40 mM N-acetylimidazole (NAI) respectively. The amount of rebound NAI-modified extrinsic 23 kDa protein was 98%, 80%, and 5% of that in the unmodified protein, respectively. These results indicate that the Tyr residues are absolutely essential to reconstitution ability. Further, the fluorescence and circular dichroism spectra among native and NAI-modified extrinsic 23 kDa proteins were similar, suggesting that the modification by NAI did not markedly influence the basic secondary structure of the native conformation. Thus, we have concluded that the tyrosine residues in the extrinsic 23 kDa protein are important for interaction with PSII membranes. In addition, we found that the structure of the extrinsic 23 kDa protein is stable in suspension (pH 4-9 or Tm 25-55 degrees C).

[Progresses in the study on light harvesting pigment protein complexes and reaction centers from purple bacteria].

This review describes the recent progress in understanding of light harvesting complexes and reaction centers from purple bacteria. Emphasis is paid on the structure of two light harvesting complexes, inner or outer, and the mechanism of the transfer of excited energy among relative pigments (Fig.1). At the same time, it is detailedly stated about the understanding of the structure of the reaction center and the transform mechanism from light energy to chemical energy, usable for life system (Fig.2).

The observation of excited-state dynamical evolution in light-harvesting complex LH2 from Rhodobacter sphaeroides 601.

With selective excitation around BChl-B800 and BChl-B850 absorption bands, we observed the evolution of excited-state dynamics in LH2 from Rhodobacter sphaeroides 601. The dynamical traces demonstrate a dominant excited-state absorption (ESA) followed concomitantly by an ultrafast transmission increase and decay with pulse-width limited time scale at 818 nm and 828 nm excitation. The ESA occurring prior to excitonic thermalization or ground-state bleach was observed at 840 nm as well. These experimental results indicate the competition between the transition from excitonic states to higher-lying excited states and interexciton relaxation, which are of physical significance for understanding excitation transfer and related mechanisms in LH2.

Photosynthesis research in the People's Republic of China.

The first research paper on photosynthesis in China was published by T.T. Li(2) in 1929. Two photosynthesis laboratories were established in Shanghai and Beijing in the 1950s and the 1960s, respectively. A photophosphorylation 'intermediate' was discovered after the energy conversion process was separated into light and dark phases in the 1960s. Since the 1980s, research has accelerated at several different levels through efforts of a large number of scientists in China.

Study on the photo-generation of superoxide radicals in Photosystem II with EPR spin trapping techniques.

Direct EPR evidence of the photo-generation of superoxide radicals (O(2) (-.)) was obtained by using a novel spin trapping probe in spinach Photosystem II (PS II) membrane fragments. The production of O(2) (-.) was detected by following the formation of 5-diethoxyphosphoryl-5-methyl-1-pyrroline-N-oxide (DEPMPO) superoxide adducts (DEPMPO-OOH). The inhibition of O(2) (-.) formation by 3-(3,4-dichlorophenyl) -1,1-dimethylurea (DCMU) and the 77 K fluorescence spectrum indicated that O(2) (-.) were generated from PS II, not from PS I. The inhibition of O(2) (-.) formation by DCMU also suggested that O(2) (-.) were generated from the Q(B)binding site, not at a site prior to DCMU blockage. The extrinsic proteins and Mn are very important to eliminate O(2) (-.), showing that the oxygen-evolving system is involved in O(2) (-.) removal rather than production.

Differentiating the orientations of photosynthetic reaction centers on Au electrodes linked by different bifunctional reagents.

The photosynthetic reaction center (RC) composite film was fabricated by self-assembled monolayers (SAMs) on the Au electrode with two different bifunctional reagents, 4-aminothiophenol (ATP) and 2-mercaptoethylamine (MEA), respectively. The square wave voltametry (SWV), bulk electrolysis and photocurrent test were employed for characterizing the composite film. The dramatic different electrochemical characteristics were observed for the two types of films, which strongly suggested an orientational difference for RC arising from the structural difference between the two bifunctional reagents. For RC-MEA film, three redox peaks which implying electron transfer (ET) between the primary donor (P) and the bacteriopheophytin (Bphe) were observed. While for RC-ATP film, two redox peaks implying ET between the nonheme iron and the primary quinone (Q(A)) were observed. The ET behavior driven by electric field also supported the result that the RC could be linked to the electrode at different sites. The site-specific immobilization approach reported here supplies a method to differentiate the protein orientation.

Studies on the Stimulating Nature of Uncouplers on the Electron Transport in BBY Particles.

BBY particles, which have kept the physicochemical property of PSII, were shown by transmission electron microscopy to possess no intact thylakoid membranes. The results of measuring 9-AA fluorescence quenching and millisecond Chl alpha delayed light emission proved that BBY particles were also unable to establish proton gradient across the membranes (deltapH) in light. Moreover, uncouplers gramicidin D and NH(4)Cl increased PSII electron transport in BBY particles only at low pH. This stimulation was more obvious around pH 6.0 than at other pH. The consistent stimulating value and pH-dependence indicated that the stimulating mechanisms of the two uncouplers are similar. From above, we infer that the uncouplers can bypass the proton transfer of localized pathway in BBY particles, stimulating the corresponding electron transport.

The Photochemical Change in Photosynthetic Reaction Center of Purple Bacterium with Pheophytin Substitution.

The reaction centers are isolated from chromatophores of Rhodobacter sphaeroides 601 by detergent LDAO, and purified by chromatography on a DEAE-52 cellulose column. In the presence of acetone and an access of free pheophytins (Phes), bacteriopheophytins (Bphes) in reaction centers are replaced by pheophytins at sites H(A) and H(B) when incubated under high temperature. The substituting amounts are about 50% and 71% Bphes in reaction centers with incubation of fifteen and sixty minutes respectively. In the absorption spectra of reaction centers containing Phes (Phe RC), the Q(X) 537 nm and Q(Y) 758 nm bands of Bphe disappeared, three distinct bands assigned to the Q(X 509/542 nm and QY) 674 nm bands of phe appeared. Compared to reaction centers in control, the photochemical activities of Phe RCs, with incubating time of fifteen and sixty minutes, drop to 78 and 71% of that in control respectively.

The Influence of NBS Modification of 241Trp on the Reconstitution of 33 kD Protein with PS and on the Recovery of Oxygen-evolving Activity.

The extrinsic 33 kD protein of photosystemII(PSII) plays an important role in the stabilizing of manganese cluster and maintaining high oxygen-evolving activity of PSII. In this research, (241)Trp, the only tryptophan in the 33 kD protein, was modified by N-Bromosuccinimide. The pH-dependence of modification suggests that this tryptophan is buried in the hydrophobic interior of the protein. The protein's capability of reconstitution to the PSII ecreased after modification, and no oxygen-evolving activity of PS was recovered after the reconstitution. Results suggest that (241)Trp of the 33 kD protein is essential for the binding of the protein to the PS and the normal oxygen-evolving activity.

[Thermodynamic and kinetic analysis of unfolding of P23k protein isolated from spinach photosystem II].

The unfolding of 23kD (P23k) protein isolated from spinach photosystem II particle was studied by high pressure and fluorescence spectroscopy. The thermal equilibrium study indicated that the protein could be totally unfolded by 180 or 160 MPa at 20 degrees C and 3 degrees C, respectively. The standard free energy and standard volume change of the protein for unfolding at 20 degrees C is 23.45 kJ/mol and -150.3 ml/mol, respectively. Kinetics study indicated that at 20 degrees C the activation volume for unfolding, delta V(u)(++), was negative (-66.2 ml/mol), meanwhile the activation volume for folding, deltaV(f)(++), was positive (84.1 ml/mol). The rate constants for folding and unfolding (K(0f), K(0u)) were 1.87 s(-1) and 1.3x10(-4) s(-1), respectively, these results provide some clues to explain why the protein is so sensitive to pressure.

The Ultrafast Energy Transfer Process in Purple Bacteria Photosynthetic Reaction Center.

The ultrafast energy transfer process, which takes place in femtosecond time range, in bacterial photosynthetic reaction center RS601 was investigated using femtosecond pump-probe technique with selective excitation. Upon 755 nmexcitation, the excited state of bacteriopheophytin H decayed to bacteriochlorophyll B with a time constant of about 130 fs, while the excited state of B transported the energy to its energy acceptor, the dimeric bacteriochlorophyll P, in about 240 fs with the 800 nm excitation. The internal conversion process between the upper and lower exciton levels of special pair P might exist upon the excitation of 850 nm pulses. In addition, from the results obtained in our experiments, the charge separation and electron transfer from P to the acceptor H was also observed via the real intermediate B within a few picoseconds.

The Characteristics of DCPIPH(2) right curved arrow MV Electron Transport in Rb. sphaeroides 601.

The absorption spectrum and fluorescence emission spectrum of RS601 were found to keep the typical characteristics of those of the purple nonsulfide bacteria Rb. sphaeroides. Under illumination, methyl viologen was reduced by RS601 chromatophores in the presence of DCPIPH(2) as the electron donor, setting up a standard noncyclic electron transport. o-phenanthroline with I(50) of 1.0 mM inhibited the DCPIPH(2) right curved arrow MV electron transport. Antimycin A did not inhibit the DCPIPH(2) right curved arrow MV electron transport and had no I(50). The results suggested that the exact site where methyl viologen accepted electron should locate between the secondary electron acceptor, Q(B), and cyt b, but not at the Q(A) binding site as indicated before. The difference of electron transport between reduced sides of reaction centers of Rb. sphaeroides and Rs. rubrum was discussed.

Comparison of Different Effects of Chloroacetates on Electron Transports in PS II and in the Reaction Center of Rb. sphaeroides 601.

Chloroacetates displayed different effects on electron transports in the photosynthetic reaction center of purple bacteria (Rb.sphaeroides 601) and in the photosystem II (PS II) of higher plants. Decays of chlorophyII a fluorescence measured after actinic flashes show that chloroacetates inhibit the electron transport from Q(A)(-) to Q(B) (Q(B)(-)) and the equilibrium between Q(A)(-)Q(B) (Q(B)(-)) and Q(A)Q(B)(-) (Q(B)(2-)), acting on electron transport as well as proton transduction. The study on PSII electron transport indicates another inhibition site of chloroacetate at the oxidation side of PSII. Chloroacetates up to 500 mM have no inhibition on the DCPIPH(2) right curved arrow MV electron transport in the RS601's chromatophores. Dissociation constants of OP and trichloroacetates from RS601 reaction center were assessed to be 1.1x10(-3) M and from Dixon curve respectively. The differences on aspects of structures and functions between RS601 reaction center and PS II were discussed.

[Different effects of illumination with xenon and sulfur lamp on growth and development of cotton plants].

Experiments were carried out with cotton (Gossgpium hirsutum cv. Xuzhou 142) plants to study the effects of illumination with xenon and sulfur lamp on development of cotton plants. The results showed that, compared with xenon lamp, illumination with sulfur lamp inhibited excessive elongation of hypocotyl via promotion of longitudinal elongation of epidermis and cortex cells, increased the numbers of branches, buds and bolls significantly. It suggested that illumination with sulfur lamp rendered cotton photomorphogenesis more favorable to yield formation.

[Differences in plant hormone level of cucumber seedlings grown under sulfur or xenon lamp].

The primary mechanism of growth difference of cucumber (Cucumis sativus L.) seedlings cultured under sulfur lamp and xenon lamp in a phytotron was investigated. Compared with cucumber seedlings grown under xenon lamp, those under sulfur lamp were shorter, and the cell number in the middle hypocotyls epidermis and cortex of them were more (Fig. 1, Table 1). Endogenous hormone analysis indicates that the content of IAA and GA(3) of seedlings under sulfur lamp were 17% and 24% lower, while ABA content was 31% higher than that under xenon lamp (Fig. 2). Based on these results, it is suggested that the growth difference between cucumber seedlings grown under sulfur lamp and under xenon lamp might be related to the control of endogenous hormones.

[Fourier transform infrared spectroscopy study on the protein secondary structure of photosystem II reaction center].

A successful study on the secondary structure of the isolated photosystem II (PSII) particles with the Fourier transform infrared spectroscopy is reported in this paper. The beta condensation effect is obviously characterized by infrared absorption spectra. The infrared spectra of both living protein and beta condensed protein samples are measured at room temperature. The amide I band in infrared spectrum is used to perform the quantitative analysis of the sample properties. The recorded spectra show the irreversible effect for the PSII particles after the 400 K heating. A rather strong change of the infrared spectra is observed due to the beta condensation of PSII protein. All the spectra are well fitted by 3-Lorentz-peak. The FTIR spectroscopy shows its effectiveness in studying the heating effect on the PSII particles.