胞内砷磷含量和比值对莱茵衣藻砷酸盐和亚砷酸盐耐性的影响
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摘要
为探索胞内砷(As)与磷(P)含量和比值与莱茵衣藻As耐性的关系,本文设置两个磷酸盐(PO_4~(3-))和系列砷酸盐(As(V))、亚砷酸盐(As(Ⅲ))浓度,处理72 h后测定莱茵衣藻生长情况和胞内As、P含量,并以培养液As浓度([As]_dis)、胞内As含量([As]_intra)、胞内砷磷比([As:P]_intra)推算半数效应浓度(EC_50),比较这3种指标对莱茵衣藻As耐性的评价效果.结果表明,随着[As]_dis的增加,莱茵衣藻[As]_intra上升,提高培养液PO_4~(3-)浓度不影响As(Ⅲ)处理下的[As]_intra,但显著降低了As(V)处理下的[As]_intra.以[As]_dis表征EC_50时,两种PO_4~(3-)水平(0.315、3.15 mg·L~(-1))下的As(Ⅲ)-EC_50(2090.3、21183.6μg·L~(-1)-As)明显高于As(V)-EC_50(162.1、2358.3μg·L~(-1)-As).基于[As]_intra的EC_50数据显示,PO_4~(3-)水平不影响As(Ⅲ)-EC_50(123.9、125.0μg·g~(-1)-As-dw),但显著影响As(V)-EC_50(7.4、58.6μg·g~(-1)-As-dw).由[As:P]_intra推算的EC_50可知,PO_4~(3-)对两种形态As毒性的影响相反,As(Ⅲ)-EC_50分别为21.1、6.1(mol/mol,As/P),As(V)-EC_50分别为1.3、3.4(mol/mol,As/P).研究结果说明,As(V)对该藻的毒性大于As(Ⅲ),莱茵衣藻对As(V)和As(Ⅲ)的耐性除了受到培养液PO_4~(3-)浓度的制约外,还受到胞内As、P含量及其比值的影响.
In order to explore the relationship between _intracellular arsenic( As) and phosphorus( P)content and their ratios with As tolerance in Chlamydomonas reinhardtii,two phosphate( PO_4~(3-))levels and a series of arsenate( As( V)) and arsenite( As( Ⅲ)) concentrations were set up,and the growth of C. reinhardtii and the contents of As and P in the C. reinhardtii cells after 72 h treatment were measured. The half maximal effective concentration values( EC_50) based on the As concentration in growth media( [As]_dis),the _intracellular arsenic content( [As]_intra) and the ratio of _intracellular arsenic to phosphorus( [As: P]_intra) were calculated. Results showed that the[As]_intraof C. reinhardtii increased with the increase of [As]_dis. Higher level of PO_4~(3-) in the culture did not affect the [As]_intraunder As( Ⅲ) treatment,but significantly decreased it under As( V)treatment. When EC_50 is characterized by [As]_dis,As( Ⅲ)-EC_50( 2090. 3,21183. 6 μg·L~(-1)-As under two PO_4~(3-) levels) were significantly higher than the As( V)-EC_50( 162.1,2358.3 μg·L~(-1)-As).The data of [As]_intraEC_50 showed that PO_4~(3-) level did not affect As( Ⅲ)-EC_50( 123.9,125.0 μg·g~(-1)-As-dw),but significantly affected As( V)-EC_50( 7. 4,58. 6 μg·g~(-1)-As-dw). From the EC_50 derived from[As: P]_intra,we found that the effect of PO_4~(3-) on the tolerance of two As species was opposite. The As( Ⅲ)-EC_50 were 21.1 and 6.1( mol/mol,As/P),while As( V)-EC_50 were 1.3 and 3.4( mol/mol,As/P). Taken together,these data indicated that As( V) was more toxic to the microalgae than As( Ⅲ). The tolerance of As( Ⅴ) and As( Ⅲ) by C. reinhardtii was not only affected by the PO_4~(3-) concentration in the culture,but also affected by _intracellular As and P content and their ratios.
In order to explore the relationship between _intracellular arsenic( As) and phosphorus( P)content and their ratios with As tolerance in Chlamydomonas reinhardtii,two phosphate( PO_4~(3-))levels and a series of arsenate( As( V)) and arsenite( As( Ⅲ)) concentrations were set up,and the growth of C. reinhardtii and the contents of As and P in the C. reinhardtii cells after 72 h treatment were measured. The half maximal effective concentration values( EC_50) based on the As concentration in growth media( [As]_dis),the _intracellular arsenic content( [As]_intra) and the ratio of _intracellular arsenic to phosphorus( [As: P]_intra) were calculated. Results showed that the[As]_intraof C. reinhardtii increased with the increase of [As]_dis. Higher level of PO_4~(3-) in the culture did not affect the [As]_intraunder As( Ⅲ) treatment,but significantly decreased it under As( V)treatment. When EC_50 is characterized by [As]_dis,As( Ⅲ)-EC_50( 2090. 3,21183. 6 μg·L~(-1)-As under two PO_4~(3-) levels) were significantly higher than the As( V)-EC_50( 162.1,2358.3 μg·L~(-1)-As).The data of [As]_intraEC_50 showed that PO_4~(3-) level did not affect As( Ⅲ)-EC_50( 123.9,125.0 μg·g~(-1)-As-dw),but significantly affected As( V)-EC_50( 7. 4,58. 6 μg·g~(-1)-As-dw). From the EC_50 derived from[As: P]_intra,we found that the effect of PO_4~(3-) on the tolerance of two As species was opposite. The As( Ⅲ)-EC_50 were 21.1 and 6.1( mol/mol,As/P),while As( V)-EC_50 were 1.3 and 3.4( mol/mol,As/P). Taken together,these data indicated that As( V) was more toxic to the microalgae than As( Ⅲ). The tolerance of As( Ⅴ) and As( Ⅲ) by C. reinhardtii was not only affected by the PO_4~(3-) concentration in the culture,but also affected by _intracellular As and P content and their ratios.
引文
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[18]丁腾达,阚啸林,吴振华,等.砷对绿藻的毒性效应及氧化还原条件的影响[J].环境化学,2016,35(5):1084-1089.DING T D,KAN X L,WU Z H,et al.Toxicity of arsenic on green alge and its effect on redox conditions[J].Environmental Chemistry,2016,35(5):1084-1089(in Chinese).
[19]MUNOZ L P,PURCHASE D,JONES H,et al.Enhanced determination of As-phytochelatin complexes in Chlorella vulgaris using focused sonication for extraction of water-soluble species[J].Analytical Methods,2014,6(3):791-797.
[20]NEARING M M,KOCH I,REIMER K J.Complementary arsenic speciation methods:A review[J].Spectrochimica Acta Part B:Atomic Spectroscopy,2014,99:150-162.
[21]WANG Y,ZHANG C H,ZHENG Y H,et al.Phytochelatin synthesis in Dunaliella salina induced by arsenite and arsenate under various phosphate regimes[J].Ecotoxicology and Environmental Safety,2017,136:150-160.
[22]BAHAR M M,MEGHARAJ M,NAIDU R.Influence of phosphate on toxicity and bioaccumulation of arsenic in a soil isolate of microalga Chlorella sp[J].Environmental Science and Pollution Research,2016,23:2663-2668.
[23]CHEN J,YOSHINAGA M,GARBINSKI L D,et al.Synergistic interaction of glyceraldehydes-3-phosphate dehydrogenase and Ars J,a novel organoarsenical efflux permease,confers arsenate resistance[J].Molecular Microbiology,2016,100(6):945-953.
[2]BAHAR M M,MEGHARAJ M,NAIDU R.Bioremediation of arsenic-contaminated water:recent advances and future prospects[J].Water,Air,&Soil Pollution,2013,224(12):1-20.
[3]WANG S,ZHAO X.On the potential of biological treatment for arsenic contaminaled soils and groundwater[J].Journal of Environmental Management,2009,90(8):2367-2367.
[4]WANG Y,WANG S,XU P,et al.Review of arsenic speciation,toxicity and metabolism in microalgae[J].Reviews in Environmental Science and Bio-Technology,2015,14:427-451.
[5]WANG N X,HUANG B,et al.Effects of nitrogen and phosphorus on arsenite accumulation,oxidation,and toxicity in Chlamydomonas reinhardtii[J].Aquatic Toxicology,2014,157:167-174.
[6]FUJIWARA S,KOBAYASHI I,HOSHINO S,et al.Isolation and characterization of arsenate-sensitive and resistant mutants of Chlamydomonas reinhardtii[J].Plant and Cell Physiology,2000,41(1):77-83.
[7]KAISE T,FUJIWARA S,TSUZUKI M,et al.Accumulation of arsenic in a unicellular alga Chlamydomonas reinhardtii[J].Applied Organometallic Chemistry,1999,13(2):107-111.
[8]YIN X,WANG L,DUAN G,et al.Characterization of arsenate transformation and identification of arsenate reductase in a green alga Chlamydomonas reinhardti[J].Journal of Environmental Sciences,2011,23(7):1186-1193.
[9]苑春刚,LE X CHRIS.砷形态分析[J].化学进展,2009,21(2/3):467-473.YUAN C G,LE X CHRIS.Arsenic speciation analysis[J].Progress in Chemistry,2009,21(2/3):467-473(in Chinese).
[10]ZHAO F,MA J,MEHARG A,et al.Arsenic uptake and metabolism in plants[J].New Phytologist,2009,181(4):777-794.
[11]LEVY J L,STAUBER J L,ADAMS M S,et al.Toxicity,biotransformation,and mode of action of arsenic in two freshwater microalgae(Chlorella sp.and Monoraphidium arcuatum)[J].Environmental Toxicology and Chemistry,2005,24(10):2630-2639.
[12]ZHANG S,RENSING C,ZHU Y G.Cyanobacteria-mediated arsenic redox dynamics is regulated by phosphate in aquatic environments[J].Environmental Science&Technology,2014,48(2):994-1000.
[13]KARADJOVA I B,SLAVEYKOVA V I,TSALEV D L,et al.The biouptake and toxicity of arsenic species on the green microalga Chlorella salina in seawater[J].Aquatic Toxicology,2008,87(4):264-271.
[14]SOMER G,NLA N.The effect of acid digestion on the recoveries of trace elements:Recommended policies for the elimination of losses[J].Turkish Journal of Chemistry,2009,30(6):745-753.
[15]ZHANG C H,WANG Y,GE Y.Determination of five arsenic species in Porphyra by microwave-assisted water extraction and high performance liquid chromatography-atomic fluorescence spectrometry[J].Analytical Letters,2013,46(10):1573-1586.
[16]BAHAR M M,MEGHARAJ M,NAIDU R.Toxicity,transformation and accumulation of inorganic arsenic species in a microalga Scenedesmus sp.isolated from soil[J].Journal of Applied Phycology,2013,25(3):913-917.
[17]WURL O,ZIMMER L,CUTTER G A.Arsenic and phosphorus biogeochemistry in the ocean:Arsenic species as proxies for P-limitation[J].Limnology and Oceanography,2013,58(2):729-740.
[18]丁腾达,阚啸林,吴振华,等.砷对绿藻的毒性效应及氧化还原条件的影响[J].环境化学,2016,35(5):1084-1089.DING T D,KAN X L,WU Z H,et al.Toxicity of arsenic on green alge and its effect on redox conditions[J].Environmental Chemistry,2016,35(5):1084-1089(in Chinese).
[19]MUNOZ L P,PURCHASE D,JONES H,et al.Enhanced determination of As-phytochelatin complexes in Chlorella vulgaris using focused sonication for extraction of water-soluble species[J].Analytical Methods,2014,6(3):791-797.
[20]NEARING M M,KOCH I,REIMER K J.Complementary arsenic speciation methods:A review[J].Spectrochimica Acta Part B:Atomic Spectroscopy,2014,99:150-162.
[21]WANG Y,ZHANG C H,ZHENG Y H,et al.Phytochelatin synthesis in Dunaliella salina induced by arsenite and arsenate under various phosphate regimes[J].Ecotoxicology and Environmental Safety,2017,136:150-160.
[22]BAHAR M M,MEGHARAJ M,NAIDU R.Influence of phosphate on toxicity and bioaccumulation of arsenic in a soil isolate of microalga Chlorella sp[J].Environmental Science and Pollution Research,2016,23:2663-2668.
[23]CHEN J,YOSHINAGA M,GARBINSKI L D,et al.Synergistic interaction of glyceraldehydes-3-phosphate dehydrogenase and Ars J,a novel organoarsenical efflux permease,confers arsenate resistance[J].Molecular Microbiology,2016,100(6):945-953.