Enhanced protection against oxidative stress in an astaxanthin-overproduction Haematococcus mutant (Chlorophyceae)

Many unicellular green algae can become yellow or red in various natural habitats due to mass accumulation of a secondary carotenoid, such as lutein, or astaxanthin. The accumulation of secondary carotenoids is generally thought to be a survival strategy of the algae under photo-oxidative stress or other adverse environmental conditions. The physiological role of the carotenoids in stress response is less well understood at the subcellular or molecular level. In this study, a stable astaxanthin overproduction mutant (MT 2877) was isolated by chemical mutagenesis of a wild type (WT) of the green microalga Haematococcus pluvialis Flotow NIES-144. MT 2877 was identical to the WT with respect to morphology, pigment composition, and growth kinetics during the early vegetative stage of the life cycle. However, it had the ability to synthesize and accumulate about twice the astaxanthin content of the WT under high light, or under high light in the presence of excess amounts of ferrous sulphate and sodium acetate. Under stress, the mutant exhibited higher photosynthetic activities than the WT, based on considerably higher chlorophyll fluorescence induction, chlorophyll autofluorescence intensities, and oxygen evolution rates. Cell mortality caused by stress was reduced by half in the mutant culture compared with the WT. Enhanced protection of the mutant against stress is attributed to its accelerated carotenogenesis and accumulation of astaxanthin. Our results suggest that MT 2877, or other astaxanthin overproduction Haematococcus mutants, may offer dual benefits, as compared with the wild type, by increasing cellular astaxanthin content while reducing cell mortality during stress-induced carotenogenesis.

Site-directed gene integration in transgenic zebrafish mediated by cre recombinase using a combination of mutant Lox sites

With current gene-transfer techniques in fish, insertion of DNA into the genome occurs randomly and in many instances at multiple sites. Associated position effects, copy number differences, and multiple gene interactions make gene expression experiments difficult to interpret and fish phenotype less predictable. To meet different fish engineering needs, we describe here a gene targeting model in zebrafish. At first, four target zebrafish lines, each harboring a single genomic lox71 target site, were generated by zebrafish transgenesis. The zygotes of transgenic zebrafish lines were coinjected with capped Cre mRNA and a knockin vector pZklox66RFP. Site-specific integration event happened from one target zebrafish line. In this line two integrant zebrafish were obtained from more than 80,000 targeted embryos (integrating efficiency about 10(-4) to 10(-5)) and confirmed to have a sole copy of the integrating DNA at the target genome site. Genomic polymerase chain reaction analysis and DNA sequencing verified the correct gene target events where lox71 and lox66 have accurately recombined into double mutant lox72 and wild-type loxP. Each integrant zebrafish chosen for analysis harbored the transgene rfp at the designated egfp concatenates. Although the Cre-mediated recombination is site specific, it is dependent on a randomly placed target site. That is, a genomic target cannot be preselected for integration based solely on its sequence. Conclusively, an rfp reporter gene was successfully inserted into the egfp target locus of zebrafish genome by Cre-lox-mediated recombination. This site-directed knockin system using the lox71/lox66 combination should be a promising gene-targeting platform serving various purposes in fish genetic engineering.

Decadal persistence of cycles in lava lake motion at Erebus volcano, Antarctica

Studies of Erebus volcano's active lava lake have shown that many of its observable properties (gas composition, surface motion and radiant heat output) exhibit cyclic behaviour with a period of ~10 min. We investigate the multi-year progression of the cycles in surface motion of the lake using an extended (but intermittent) dataset of thermal infrared images collected by the Mount Erebus Volcano Observatory between 2004 and 2011. Cycles with a period of ~5-18 min are found to be a persistent feature of the lake's behaviour and no obvious long-term change is observed despite variations in lake level and surface area. The times at which gas bubbles arrive at the lake's surface are found to be random with respect to the phase of the motion cycles, suggesting that the remarkable behaviour of the lake is governed by magma exchange rather than an intermittent flux of gases from the underlying magma reservoir. © 2014 The Authors.

Compensatory growth and food consumption in gibel carp, Carassius auratus gibelio, and Chinese longsnout catfish, Leiocassis longirostris, experiencing cycles of feed deprivation and re-feeding

The compensatory responses of juvenile gibel carp and Chinese longsnout catfish to four cycles of 1 part of a study designed to determine feeding regimes that would maximise growth rates. Both species showed compensatory growth in the re-feeding periods. The compensation was not sufficient for the deprived fish to match the growth trajectories of controls fed to satiation daily. The compensatory growth response was more clearly defined in the later cycles. The deprived fish showed hyperphagia during the 2-week periods of re-feeding and the hyperphagic response was clearer in the later cycles. The hyperphagia tended to persist for both weeks of the re-feeding period. The gibel carp showed no difference in gross growth efficiency between deprived and control fish. In the catfish, the gross growth efficiency of the deprived fish was marginally higher than that of control fish, but the efficiency varied erratically from week to week. Over the experiment, the deprived fish achieved growth rates 75-80% of those shown by control fish, although fed at a frequency of 66%. There was no evidence of growth over-compensation with the deprivation-re-feeding protocol used in this study. (C) 2004 Elsevier B.V. All rights reserved.

Effect of cycles of feed deprivation on growth and food consumption of immature three-spined sticklebacks and European minnows

Individual juvenile three-spined sticklebacks Gasterosteus aculeatus and European minnow Phoxinus phoxinus, from sympatric populations, were subjected to four cycles of I week of food deprivation and 2 weeks of ad libitum feeding. Mean specific growth rate during the weeks of deprivation was negative and did not differ between species. The three-spined stickleback showed sufficient growth compensation to recover to the growth trajectory shown by control fish daily fed ad libitum. The compensation was generated by hyperphagia during the re-feeding periods, and in the last two periods of re-feeding, the gross growth efficiencies of deprived three-spined sticklebacks were greater than in control fish. The expression of the compensatory changes in growth and food consumption became clearer over the successive periods of re-feeding. The European minnow developed only a weak compensatory growth response and the mass trajectory of the deprived fish deviated more and more from the control trajectory During re-feeding periods, there were no significant differences in food consumption or gross growth efficiency between control and deprived European minnows. The differences between the two species are discussed in terms of the possible costs of compensatory growth, the control of growth and differences in feeding biology (C) 2003 The Fisheries Society of the British Isles.

Construction and analysis of a high-CO2-requiring mutant of the cyanobacterium Anabaena sp strain PCC7120

A mutant of Anabaena sp. strain PCC7120 requiring high CO2 was generated using Tn5 mutagenesis. This is the first data for a filamentous cyanobacterium. The mutant was capable of growing at 5% CO2, but incapable of growing at air levels of CO2. Southern hybridization analysis indicated that the Anabaena genome was inserted by the transposon at one site. The apparent photosynthetic affinity of the mutant to external dissolved inorganic carbon (DIC) was about 300 times lower that of the wild type (WT), and the medium alkalization rate as well as the carboxysomal carbonic anhydrase activity of the mutant was also lower than those of the WT. When the mutant was transferred from the culture medium bubbled with 5% CO2 to higher DIC (8.4% CO2) or 1% CO2, it showed similar responses to the WT. However, aberrant carboxysomes were found in the mutant cells through ultrastructural analysis, indicating it was most probably the wrong organization of the carboxysomes that eventually led to the inefficient operation of carboxysomal carbonic anhydrase and the subsequent defectiveness of the mutant in utilizing DIC.

Selection and ultrastructural observation of a high-CO2-requiring mutant of cyanobacterium Synechococcus sp PCC7942

A high-CO2-requiring mutant of Synechococcus sp. PCC7942 las been isolated after chemical mutagenesis of ethyl methane sulphonate (EMS). It was able to grow at 4% CO2, but not under ambient CO2. The initial screening of the mutant showed that the genetic reversion rate was about 10(-7) and death occurred 2 -3 days after being transferred from 4% CO2 to the ambient air. Its photosynthetic dependence on external dissolved inorganic carbon was higher than that of the wild type cells, but its carbonic anhydrase activity was comparatively low. In the ultrastructural level, various types of aberrant carboxysomes appeared in the mutant cells: rod-shaped carboxysomes, irregular carboxysomes and the "empty-inclusion carboxysomes" with increasing number of glycogen granules surrounding the thylakoids. All these alterations indicated that the mutant was defective in utilizing the external CO2. The induction of carboxysomes by lower levels of CO2 and the biogenesis of carboxysomes are herein discussed.

Research on the ultrafast fluorescence property of thylakoid membranes of the wild-type and mutant rice

A high yielding rice variety mutant (Oryza sativa L., Zhenhui 249) with low chlorophyll b (Chl b) has been discovered in natural fields. It has a quality character controlled by a pair of recessive genes (nuclear gene). The partial loss of Chl b in content affects the efficiency of light harvest in a light harvest complex (LHC), thus producing the difference of the exciting energy transfer and the efficiency of photochemistry conversion between the mutant and wild-type rice in photosynthetic unit. The efficiency of utilizing light energy is higher in the mutant than that in the wildtype rice relatively. For further discussion of the above-mentioned difference and learning about the mechanism of the increase in the photochemical efficiency of the mutant, the pico-second resolution fluorescence spectrum measurement with delay-frame-scanning single photon counting technique is adopted. Thylakoid membranes of the mutant and the wild-type rice are excited by an Ar+ laser with a pulse width of 120 ps, repetition rate of 4 MHz and wavelength of 514 nm. Compared with the time and spectrum property of exciting fluorescence, conclusions of those ultrafast dynamic experiments are: 1) The speeds of the exciting energy transferred in photo-system I are faster than that in photo-system II in both samples. 2) The speeds of the exciting energy transfer of mutant sample are faster than those of the wildtype. This might be one of the major reasons why the efficiency of photosynthesis is higher in mutant than that in the wild-type rice.

Experimental study on polarization holographic storage in genetic mutant BR-D96N film

Recording with both parallel and orthogonal linearly polarized lights, polarization holographic storage in genetic mutant BR-D96N film is reported with both transmission type geometry and reflection type geometry. Polarization properties of diffraction light and scattering light are discussed for two different cases, parallel polarization recording and orthogonal polarization recording. It shows that, compared with recording with parallel polarization lights, orthogonal polarization holography can separate the diffraction light from the scattering noise, therefore improving the signal-to-noise ratio. It also shows that, compared with reconstruction with reference light, reconstruction with phase conjugated wave of the reference light can improve the signal-to-noise ratio of the reconstructed diffraction image, and also the wave-front aberration of the object light introduced by irregular phase object in the optical pass-way can also be corrected effectively, which ensures that the reconstructed diffraction image has a better fidelity. The preliminary angle-multiplexed volume holographic storage multiplexed by transmission type geometry and reflection type geometry is demonstrated in the BR-D96N film. Experiment shows that there is no cross-talk between the two pages of images except for some scattering noises.

CFD analysis of thermodynamic cycles in a pulse tube refrigerator

The objectives of this paper are to study the thermodynamic cycles in an inertance tube pulse tube refrigerator (ITPTR) by means of CFD method The simulation results show that gas parcels working in different parts of ITPTR undergo different thermodynamic cycles The net effects of those thermodynamic cycles are pumping heat from the low temperature part to the high temperature part of the system The simulation results also show that under different frequencies of piston movement the gas parcels working in the same part of the system will undergo the same type of thermodynamic cycles The simulated thermal cycles are compared with those thermodynamic analysis results from a reference Comparisons show that both CFD simulations and theoretical analysis predict the same type of thermal cycles at the same location However only CFD simulation can give the quantitative results while the thermodynamic analysis is still remaining in quality (C) 2010 Elsevier Ltd All rights reserved