Enhancement of nitrogen and phosphorus removal from eutrophic water by economic plant annual ryegrass (Lolium multiflorum) with ion implantation.

Severe eutrophication of surface water has been a major problem of increasing environmental concern worldwide. In the present study, economic plant annual ryegrass (Lolium multiflorum) was grown in floating mats as an economic plant-based treatment system to evaluate its potential after ion implantation for removing nutrients in simulated eutrophic water. The specific weight growth rate of L. multiflorum with ion implantation was significantly greater than that of the control, and the peroxidase, nitrate reductase, and acid phosphatase activities of the irradiated L. multiflorum were found to be greater than those plants without ion implantation. Higher total nitrogen (TN) and total phosphorus (TP) removal efficiencies were obtained for the L. multiflorum irradiated with 25 keV 5.2 × 10(16) N(+) ions/cm(2) and 30 keV 4.16 × 10(16) N(+) ions/cm(2), respectively (p < 0.05). Furthermore, the nitrogen and phosphorus contents in the plant biomass with ion implantation were also greater than those in the control and were positively correlated with TN and TP supplied. L. multiflorum itself was directly responsible for 39-49 and 47-58 % of the overall N and P removal in the experiment, respectively. The research results suggested that ion implantation could become a promising approach for increasing phytoremediation efficiency of nutrients from eutrophic water by L. multiflorum.

L(+)-lactic acid production by co-fermentation of glucose and xylose with Rhizopus oryzae obtained by low-energy ion beam irradiation.

Low-energy ion beam irradiation (10-200 keV) has been proved to have a wide range of biological effects in recent years. When Rhizopus oryzae PW352 was irradiated with a 15-keV low-energy ion beam an L(+)-lactic acid high-yield mutant, RQ4015, was obtained. When 150 g/l glucose was used as the sole carbon source, L(+)-lactic acid of RQ4015 reached 121 g/l after 36 h shake-flask cultivation. However, the highest lactic acid concentration 74 g/l was obtained when 100 g/l xylose was present in the medium as the sole carbon source. When mixed xylose (25 g/l) and glucose (75 g/l) were present in a bubble column, L(+)-lactic acid production of RQ4015 reached 83 g. A high mutation rate and a wide mutation spectrum of low-energy ion implantation were observed in the experiment, suggesting that ion implantation can be a highly efficient mutagenic means for microorganism breeding in many commercial applications.

Enhanced nitrogen and phosphorus removal from eutrophic lake water by Ipomoea aquatica with low-energy ion implantation.

Ipomoea aquatica with low-energy N+ ion implantation was used for the removal of both nitrogen and phosphorus from the eutrophic Chaohu Lake, China. The biomass growth, nitrate reductase and peroxidase activities of the implanted I. aquatica were found to be higher than those of I. aquatica without ion implantation. Higher NO3-N and PO4-P removal efficiencies were obtained for the I. aquatica irradiation at 25 keV, 3.9 x 10(16) N+ ions/cm(2) and 20 keV 5.2 x 10(16) N+ ions/cm(2), respectively (p < 0.05). Moreover, the nitrogen and phosphorus contents in the plant biomass with ion implantation were also greater than those of the controls. I. aquatica with ion implantation was directly responsible for 51-68% N removal and 54-71% P removal in the three experiments. The results further confirm that the ion implantation could enhance the growth potential of I. aquatica in real eutrophic water and increase its nutrient removal efficiency. Thus, the low-energy ion implantation for aquatic plants could be considered as an approach for in situ phytoremediation and bioremediation of eutrophic waters.

Enhanced enzymatic saccharification of rice straw by microwave pretreatment.

In this study, Box-Behnken design and response surface methodology were employed to plan experiments and optimize the microwave pretreatment of rice straw. Experimental results show that microwave intensity (MI), irradiation time (IT) and substrate concentration (SC) were main factors governing the enzymatic saccharification of rice straw. The maximal efficiencies of cellulose, hemicellulose and total saccharification were respectively increased by 30.6%, 43.3% and 30.3% under the optimal conditions of MI 680 W, IT 24 min and SC 75 g/L. The chemical composition analysis of straw further confirmed that microwave pretreatment could disrupt the silicified waxy surface, break down the lignin-hemicellulose complex and partially remove silicon and lignin.

A novel approach to microbial breeding--low-energy ion implantation.

Low-energy ions exist widely in the natural world. People had neglected the interaction between low-energy ions and material; it was even more out of the question to study the relation of low-energy ions and the complicated organism until the biological effects of low-energy ion implantation were discovered in 1989. Nowadays, the value of low-energy ion beam implantation, as a new breeding way, has drawn extensive attention of biologists and breeding experts. In this review, the understanding and utilization of microbial breeding by low-energy ion beam irradiation is summarized, including the characteristics of an ion beam bioengineering facility, present status of the technology of low-energy ions for microbial breeding, and new insights into microbial biotechnology.

Nitrogen removal from eutrophic water by floating-bed-grown water spinach (Ipomoea aquatica Forsk.) with ion implantation.

The aim of this study was to investigate the use of water spinach (Ipomoea aquatica Forsk.) with N(+) ion-beam implantation for removal of nutrient species from eutrophic water. The mutated water spinach was grown on floating beds, and growth chambers were used to examine the growth of three cultivars of water spinach with ion implantation for 14 days in simulated eutrophic water at both high and low nitrogen levels. The specific weight growth rates of three cultivars of water spinach with ion implantation were significantly higher than the control, and their NO(3)-N and NH(4)-N removal efficiencies were also greater than those of the control. Furthermore, compared with the control, the nitrogen contents in the plant biomass with ion implantation were higher as well.

Mutational spectrum of the lacI gene in Escherichia coli K12 induced by low-energy ion beam.

The mutational spectrum of the genomic lacI gene induced by low-energy nitrogen ion irradiation in wild type Escherichia coli strain W3110 were compared with the spontaneous and the vacuum controls. The mutant frequency of irradiated group was dose-dependent and reached 26.3 x 10(-6) at dose of 31.2 x 10(14) ions/cm2, which was about 18-fold over the background (1.5 x 10(-6)) and 10-fold over the vacuum controls (2.6 x 10(-6)). This result indicated that the low-energy ion irradiation was one of many effective mutagens, though the vacuum condition of low-energy ions contributed some low-level gene mutations. It was found that the difference between the spontaneous and the vacuum control was the increases of base-pair substitutions in the vacuum control group. The spectra of irradiated group were quite similar to that of oxygen free-radical induced in the same strain, suggesting free-radicals and other adducts generated by low-energy ions might play an important role in the mutagenesis in vivo. When the spontaneous and the vacuum control group were compared, base-pair substitutions, deletions and additions of the irradiated group were significantly increased, and the +TGGC or -TGGC at hot spot was decreased from 82 to 48%. But the remarkable increase in absolute MF of the +TGGC or -TGGC at hot spot in the irradiated group suggested that low-energy ions did induce the mutations of this type. The spectra of our irradiated group had relative low-level base-pair substitutions, high-level +/-TGGC and high proportion additions than those of gamma-radiation induced, implying there were some different effects or processes between them.

A novel approach for improving the productivity of ubiquinone-10 producing strain by low-energy ion beam irradiation.

Agrobacterium tumefaciens ATCC4452 cells were irradiated by nitrogen ion beam, a new mutagen, with energy of 10 keV and fluence ranging from 2.6x10(14) ions/cm2 to 6.5x10(15) ions/cm2. A similar "saddle shape" survival curve due to ion beam irradiation appeared again in this study. Some mutants with high yield of ubiquinone-10 were induced by ion implantation. High mutation rate and wide mutation spectrum were also observed in the experiment. These results suggested that the mutagenic effect of such low-energy ion influx into bacterium cells could result from multiple processes involving direct collision of particles with cytoplasm, nucleolus, and cascade atomic and molecular reactions due to plentiful primary and secondary particles.

[Study on breeding of probiotic Aspergillus nigre by N+ implantation and fermentation condition of its mutant].

Using Aspergillus niger AN01 as original strain, mutated strain Aspergillus niger AN03 was obtained by N+ implantation. The results showed that activities of acidic protease, cellulase and pectinase of Aspergillus niger were raised from 71.6U/g, 141.7U/g and 264.8U/g to 996.5U/g, 940.4U/g and 906.5U/g respectively. Characteristics of enzymes production in the mutant Aspergillus niger AN03 kept stabile by 5 times subculture. In addition, the optimum conditions for enzymes production of Aspergillus niger AN03 were investigated. The optimum components of medium consisted of bran 105.0g, corn straw 105.0g, bean cake 105.0g, NH4Cl 6.0g, H2O 1000mL. The optimum fermentation condition was incubated for 4d in the condition of pH 5.0 and temperature 30 degrees C.

[CD59 mutation and DNA oxidative damage in A(L) cells induced by crocidolite fibers].

OBJECTIVE: To determine the effects of buthionine sulfoximine (BSO) and free radical scavenger, dimethyl sulfoxide (DMSO), on mutation frequency and the formation of 8-hydroxydeoxyganosine (8-OHdG) induced by crocidolite fibers in human-hamster hybrid (A(L)) cells. METHODS: The cytotoxicity and mutagenicity were determined by the formation of colonies. 8-OHdG was examined by immunoperoxidase staining. Non-protein sulfhydryl (NPSH) compound was assayed by modified Tietze's method. RESULTS: The level of NPSH in A(L) cell pretreated with 25 micro mol/L of BSO was decreased to 2 nmol/10(7) cells, only 5% of the control after 24 h. The mutation frequency of CD59 gene of A(L) cell in crocidolite alone treated group was 208 +/- 18 while that in BSO pretreated group (397 +/- 55) was about twice the former (P < 0.05). The mutation frequency of CD59 gene in the group treated with crocidolite and in the presence of DMSO (57 +/- 8) was 72.6% less than that in crocidolite alone treated group. Crocidolite fibers induced a dose-effect relationship in the formation of 8-OHdG in A(L) cells (y = 150 + 20x, r = 0.9621). The level of 8-OHdG in cells was 289 +/- 6 at the dose of 6 micro g/cm(2) crocidolite, which was about twice the control group (137 +/- 9). In the presence of DMSO, 8-OHdG level decreased to 170 +/- 3 at the same dose of crocidolite. CONCLUSION: Free radicals are the important inducer of mutagenesis and DNA damage in A(L) cells caused by crocidolite, which has dose-effect relationship.

Comparison of base substitutions in response to nitrogen ion implantation and 60Co-gamma ray irradiation in Escherichia coli

To identify the specificity of base substitutions, a novel experimental system was established based on rifampicin-resistant (Rif r) mutant screening and sequencing of the defined region of the rpoB gene in E. coli. We focused on comparing mutational spectra of base substitutions induced by either low energy nitrogen ion beam implantation or 60Co-gamma rays. The most significant difference in the frequency of specific kinds of mutations induced by low energy nitrogen ion beam was that CG -> TA transitions were significantly increased from 32 to 46, AT -> TA transversions were doubled from 7 to 15 in 50 mutants, respectively. The preferential base substitutions induced by nitrogen ion beam implantation were CG -> TA transitions, AT -> GC transitions, AT -> TA transversions, which account for 92.13 percent (82/89) of the total. The mutations induced by 60Co-gamma rays were preferentially GC -> AT and AT -> GC transitions, which totaled 84.31 percent (43/51).

[Assessment of the Escherichia coli Tat protein translocation system with fluorescent proteins].

The possibility of using fluorescent proteins as probes to study the twin-arginine translocation (Tat) system was assessed in Escherichia coli. When fused to the twin-arginine signal peptide of trimethylamine N-oxide reductase, the DsRed2 red fluorescent protein from the Discosoma sp. was successfully synthesized and folded in E. coli cells. However, RR-DsRed2 aggregated inside the cells. Therefore, although DsRed2 has been engineered from DsRed for faster maturation and lower non-specific aggregation, it is still not compatible with Tat-dependent translocation. In contrast, the jellyfish green fluorescent protein (GFP) was efficiently exported into periplasm even when the RR motif was changed to KR or RK. These results show that GFP can be used as an efficient reporter protein to study Tat system, but DsRed2 is not suitable for such purpose because of its aggregation property. In addition, when the protein concentration was similar, the fluorescence intensity of KR-GFP and RK-GFP decreased compared with RR-GFP, which would suggest that the twin-arginine signal peptide is not only essential for mediating protein translocation, but also important for the folding of down-stream protein.

[Progress of chlamydomonas as a model organism].

The unicellular alga Chlamydomonas offers a simple life cycle, easy culture and isolation of series of mutants, established the techniques and tool kit for molecular genetics and genetics analysis. It is now becoming the model organism for studies on photosynthesis in plant, flagellar assembly and function, cell cycle and circadian rhythms, signal transduction, light perception and cell recognition. It is summarized the progress of study on Chlamydomonas as a model organism in this paper.

[Molecular basis of plant nuclear male sterility].

With the cloning of nuclear male fertility genes, learning of nuclear male fertility not only limited in genetic analysis and cell biology, but also advanced in molecular basis of male nuclear sterility and postulated in possible mechanisms of nuclear male sterility. The objectives of this paper are to review the related progress of plant nuclear male sterility.

Kinetics of Ionizing Irradiation-induced DNA Strand Breaks.

By measuring the dose effect of ionizing irradiation-induced single- and double-strand breaks (SSB and DSB) of pBR322, the responses of the yields of SSB and DSB to DNA concentrations were obtained. It was found that, in the presence of mannitol, the reciprocals of G(SSB) and G(alphaDSB) had a linear relationship to the reciprocal of DNA concentration so that the competition reaction of DNA and mannitol to.OH radicals could be described by a second-order kinetics. Moreover, the rate coefficients and the efficiencies of the.OH radical inducing SSB and alphaDSB were deduced.

Mechanisms of Formation of DNA Strand Breaks Induced by gamma-Rays.

Dose responses of DNA single- and double-strand breaks (SSB and DSB) of pBR322 irradiated with gamma-rays were measured by gel electrophoresis scanning and the production process of SSB was tested. Results showed that the formation of DSB was related to the free radicals scavenging capacity sigma of mannitol dissolved in DNA solution. The ratio of alphaDSB to betaDSB non-linearly increased with increasing sigma. On the other hand alphaDSB was induced by the single radical transfer mechanism as sigma<10(8)s(-1) it could however be formed by the LMDS mechanism as sigma>2x10(8)s(-1). Moreover the distance for two SSBs forming a DSB in complementary strands was not obviously influenced by the scavenging capacity.

Reverse Oxygen Effect on gamma-Irradiation Induced Single Strand Breaks of Plasmid DNA.

Oxygen effect on gamma-irradiation induced single strand breaks (SSB) of plasmid DNA was measured by the method of gel scanning. It was found that, in the presence of mannitol, the yield of SSB in N(2)-saturated DNA solution was larger than that in air-saturated solution so that the oxygen enhancement ratio (OER) of SSB became smaller than 1, which indicates the appearance of reverse oxygen effect of DNA SSB. At the same time, G(SSB) and its OER decreased with increasing concentration of mannitol.