Glutathione2Related Enzymes of Scenedesmus quadrica uda

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1 ENVIRONMENTAL SCIENCE Vol 30,No 1 Jan,2009, 3, (, ) :, ( ) 2S2 ( GST) ( GPx),, [ Pb(NO 3 ) 2 ] (HgCl 2 ) 96h EC mgπl mgπl, AI 01009, 12 h, 70 %, ; GST,, % ; GPx, % GST GPx : ; ; ; ; ; GST; GPx :X173 :A : (2009) Individual and Joint Stress of Lead and Mercury on Glutathione2Related Enzymes of Scenedesmus quadrica uda LI Yan, ZHU Lin, LIU Shuo Growth, Glutathione and (Tianjin Key Laboratory of Environmental Remediation and Pollution Control, College of Environmental Science and Engineering, Nankai University,Tianjin ,China) Abstract :To understand the toxicity mechanisms of mixed heavy metals on aquatic plant,indicators of algea growth rate,content of reduced glutathione ( ),activities of glutathione S2transferase ( GST) and glutathione peroxidase ( GPx) of green algae, Scenedesmus quadricauda were measured to analyze the individual and joint toxic effects of lead and mercury The results show that the 96h EC 50 of algae growth inhibition by lead [ Pb(NO 3 ) 2 ] and mercury (HgCl 2 ) are mgπl and mgπl respectively After 12 h individual and joint lead and mercury exposure,the content of in alga cells is decreased to about 70 % of the level of the control,and keeps a steady level with the increase of the exposure concentration The GST activities are increased to a peak in lower concentration groups and then decrease with the increase of the exposure concentration Indeed,the higher concentration of lead and mercury combined2poisoning can inhibit the activities of GST significantly, with % inhibitory rate The activity of GPx is almost suppressed continuously with the increase of the exposure concentration,and the lowest activity is only % of the control The toxic action of the mixture of Pb and Hg on growth inhibition, content,activities of GST and activities of GPx for Scenedesmus quadricauda are addition Key words : Scenedesmus quadricauda ; lead ; mercury ; joint toxicity ; ; GST; GPx,, ( ) ( GPx) 2S2 ( GST), ( GR), [1,2 ] ROS, GPx (ROS), GST,, (LPX),, ROS, : ; : : (973) (2004CB418504) : (1977 ),,,,E2mail nankai edu cn 3,E2mail edu cn [3 ]

2 1 : 249 I [4 ] A = A C - A T 100 (1) A C,, I A ; A C ; A T, [5,6 ], (1), 96 h EC 50,,,, 0 ( ), mgπl Pb (NO 3 ) 2 0 ( [7,8 ] [9,10 ], ) mgπl HgCl 2, [11 ],, 3 (2), 96 h EC 50, 015 EC 50 ( ) EC 50 ( ) = ( ) (TU ), Pb (NO 3 ) 2 HgCl 2 ( Scenedesmus quadricauda) 1 3, 96 h EC [ Pb (NO 3 ) 50 2 ] (AI) [13 ] Pb (NO (HgCl 2 ), 1 gπl Pb (NO 3 ) 3 ) 2 HgCl gπl, : HgCl 2 OECD201 [12 ] M = (2) i = 1 EC 50 i M 1,AI = (3) M M > 1,AI = M ( - 1) (4) 250 ml,, M ; c i EC 50 i 100 ml, ΠmL ; EC 50 i i EC 50 ;AI, 25 1, lx, L D = 12 h 12 h, 6 h 1,, [12 OECD201 ], 96 h 24 h 1, XB2K225, Olympus (TU ),, (1) ( I A ), 72 h,, 96 h EC h EC 50 n 1 Pb( NO 3 ) 2 HgCl 2 Πmg Table 1 Concentration of Pb(NO 3 ) 2 and HgCl 2 in joint c i treatment groupπmg L - 1 TU Pb(NO 3 ) 2 HgCl L - 1

3 = 1 ( TU) 2,, 12 h, TU, (1) rπmin 2 1) ΠTU Table 2 Concentration of Pb(NO 3 ) 2 and HgCl 2 ΠTU Pb(NO 3 ) (0 0) 013 ( ) 014 ( ) 016 ( ) 017 ( ) 019 ( ) HgCl (0 0) 012 ( ) 014 ( ) 015 ( ) 016 ( ) 018 ( ) Pb(NO 3 ) 2 + HgCl ( ) 1) Πmg L ( ) 013 ( ) 015 ( ) 017 ( ) 019 ( ) 10 min, 2 3, (ANOVA) LSD,, p molπl (ph 714), 2 min 4, rπmin 10 min, 211 GPx GST (1) ( I A ), (2) GPx GST (22 ) (DTNB) [14 ], GPx Hafemen [15 ],GST Habig [16 ] 96 h EC 50, 3, 96 h EC mgπl, (3) Bradford [17 ] 4 5 HgCl 2 Pb (NO 3 ) 2, (BSA), 96 h EC 50, gπml, M = AI = (Means SD), SPSS 1310 GST GPx 3 96 h EC 50 Table 3 96 h EC 50 of single and joint effects of Pb(NO 3 ) 2 and HgCl 2 on Scenedesmus quadricauda n r 96 h EC % Pb(NO 3 ) 2 I A = c mgπl HgCl 2 I A = c mgπl Pb(NO 3 ) 2 + HgCl 2 I A = c c TU ( % %) ( % 12 h, 7147 %), ( p 1, 0105) HgCl 2, 016 TU,,, ( p 0105) % 6180 %(018 TU, p 0105), % % (012 TU, p 0105) 016 TU(63106 % 0192 %, p 0105), 014, TU

4 1 : 251 ( p 0105) (68178 % 4167 %,013 TU, p 0105),, (66147 % 1137 %) (75127 % 6138 %), TU ( p 0105) 2 GST Fig 2 Toxic effects of single and joint Pb(NO 3 ) 2 and HgCl 2 on GST of Scendesmus quadricauda 1 Fig 1 Toxic effects of single and joint Pb(NO 3 ) 2 and HgCl 2 on of Scenedesmus quadricauda %(019 TU, p 0105) % 9168 % ( GST TU, p 0105) % 8193 % ( 017 TU, p 0105) GST 2,,GST HgCl 2, 014 TU % 2180 %( p 0105), % % (018 TU, p 0105), GST,, ( p 0105) GST 016 TU, % 3168 % ( p 0105),, GST ( % %, 019 TU, p 0105), GST, 011 TU, GPx ( p 0105), GPx % 3 GPx Fig 3 Toxic effects of single and joint Pb(NO 3 ) 2 and HgCl 2 on GPx of Scenedesmus quadricauda % 7167 %( p 0105) GST 3, TU,,GST % 3128 %( p, GST 0105) GST mrna [ GPx ] ROS GPx, GST mrna, 3, GST GST GST

5 (48 h),,, (30 d), [20 ],,, GST,, ROS 12 h,,,, GST,,, GST,,, 1975, GST Hutchinson [25 ], GST,,,, AI = 0,, (addition) ; AI < 0, GST,, (antagonism) ; AI > 0,,, (synergism),,, Fern ndez [26 ],,, M ,,, M, = ,AI = ,,,,ROS GST, GPx GST GPx,,, GPx GST,, GST, ;, GST GPx, GPx GST, ROS, GPx, GPx,,, GPx, GST GPx, [21,22 ], GPx, [23 ], GPx, GST GPx Okamoto [24 ],

6 1 : 253,,,, GST GPx,, GST GPx 4 (1) 96 h EC mgπl mgπl, (2),, (3) GST, (4) GPx ROS (5) GST GPx : [ 1 ] Assche F V,Clijsters H Effects of metals on enzyme activity in plants [J ] Plant,Cell and Environ,1990,13 (3) : [ 2 ] Bidar G,Garcon G,Pruvot C, et al Behavior of Trifolium repens and Lolium perenne growing in a heavy metal contaminated field Plant metal concentration and phytotoxicity [J ] Environ Pollut,2007,147 (3) : [ 3 ] Mittler R Oxidative stress, antioxidants and stress tolerance [ J ] Trends Plant Sci,2002,9(7) : [ 8 ],,, [J ],1999,23(1) :1210 [ 9 ] [J ],2001,25 (3) :87290 [10 ] Fang J, Wang K X, Tang J L, et al Copper, lead,zinc, cadmium, mercury and arsenic in marine products of commerce from Zhejiang coastal area, China,May 1998 [J ] Bull Environ Contam Toxicol, 2004,73 (3) : [11 ], [J ],2000,20 (5) : [12 ] OECD guidelines for the testing of chemicals 201,Freshwater alga and cyanobacteria,growth inhibition test [ S] [13 ] Making L L An Approach for Additive Toxicity of Chemical Mixtures [A] In :Mayor L L, Hamelink J L (eds) Aquatic Toxicology and Hazard Evaluation ; ASTM STP 634 [ C ] Philadelphia, American Society for Testing and Material, [14 ] Ellman G L Tissue sulfhydryl group [J ] Arch Biochem Biophys, 1959,82 :70277 [15 ] Hafeman D G,Sunde R A,Hoekstra W G Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat [J ] J Nutr, 1974,104 (5) : [16 ] Habig W H,Pabst M J,Jakoby W B Glutathione S2transferases The first enzymatic step in mercapturic acid formation [J ] J Biol Chem, 1974,249 (22) : [17 ] Bradford M M A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein2dye binding [J ] Anal Biochem,1976,72 (7) : [18 ] Marrs K A The functions and regulation of glutathione S2transferases in plants[j ] Annu Rev Plant Mol Biol,1996,47 : [19 ] Conklin P L, Last R L Differential accumulation of antioxidant mrnas in Arabidopsis thaliana exposed to ozone [J ] Plant Physiol, 1995,109 (1) : [20 ] Edwards R,Dixon D P, Walbot V Plant glutathione S2transferases : enzymes with multiple functions in sickness and in health[j ] Trends Plant Sci,2000,5(5) : [21 ],, Hg 2 + [J ],2007,27(7) : [22 ],,, Cd [J ],2007,26(3) : [23 ] Ding Y,Miao J L,Li G Y Effect of Cd on and 2related [ 4 ] Cantoni O, Christie N T, Swann A, et al Mechanism of HgCl 2 cytotoxicity in cultured mammalian cells [J ] Mol Pharmacol,1984, 26(2) : enzymes of Chlamydomonas sp ICE2L existing in Antarctic ice [J ] J Environ Sci2China,2005,17(4) : [24 ] Okamoto O K,Pinto E,Latorre L R, et al Antioxidant modulation in [ 5 ],,, response to metal2induced oxidative stress in algal chloroplasts [J ] [J ],2006, Arch Environ Contam Toxicol,2001,40 (1) : (2) :53255 [ 6 ] Fatima R A,Ahmad M Certain antioxidant enzymes of Allium cepa as biomarkers for the detection of toxic heavy metals in wastewater [J ] Sci Total Environ,2005,346 (123) : [ 7 ] Qu C H, Yan R E Chemical composition and factors controlling [25 ] Hutchinson T C, Stockes P M Heavy Metal Toxicity and Algal Bioassays [ M ] Philadelphia : American Society for Testing and Materials, [26 ] Fern ndez N,Beiras R Combined toxicity of dissolved mercury with copper,lead and cadmium on embryogenesis and early larval growth of suspended matter in three major Chinese rivers [ J ] Sci Total the Paracentrotus lividus Sea2Urchin [ J ] Ecotoxicology, 2001, 10 Environ,1990,97Π98 : (5) :

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