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1 31 6 Vol. 31,No ACTA HYDROBIOLOGICA SINICA Nov., DMSO ASA ROS 1, (1., ; 2., ; 31, ) : 2 g/ ml 2RR (MC2RR) 2 g/ ml MC2RR % (DMSO) 2 g/ ml MC2RR + 2 mmol/ L (ASA), (ROS),,MC2RR ROS (MDA) ASA (SOD) (POD), ( GSH) DMSO ASA, ROS MDA,ASA GSH SOD POD,, ROS, ASA DMSO MC2RR, : 2RR ; ; ; ;ASA ;DMSO :Q26 :A : (2007) ,,, ( Microcystis aeruginosa) ( Anabaena) ( Oscillatoria) [1 ] 60 (MC), 7,MC 1 2A ( PP1 PP2A), PP1 PP2A [2 ],,MC [3 ] Pflugmacher MC2LR [4 ] ; MC2RR ( ROS) [5 ] ; MC2RR,ROS [6 ] Pinho E, [7 ] ; Οegura ( GSH) MC Hep G2 ROS DNA [8 ], (ASA), H 2 O 2, (DMSO), ASA DMSO, ROS,, MC : ; : : CB ; AA ; : (1982 ),, ; ;E2mail ac. cn :, E2mail ac. cn

2 ( Nicotiana1 tabacum L1cv1Bright Yellow 2) MS (MS,015 mg/ L,113 mg/ L B1,200 mg/ L KH 2 PO 4 3 %(W/ V),012 mg/ L 2,42 D 011 mg/ L ), ph 518, 130 r/ min, (25 1),24h 1. 2 MC2RR, 5 % 3h, 8000 r/ min 30min, 2, 3, C18, 90 %,, HPLC, :2 g/ ml MC2RR, : MS MC2 RR, 2 g/ ml 2 :015 % DMSO + 2 g/ ml MC2RR, : MS MC2RR DMSO, 2 g/ ml 015 % 3 :2 mmol/ LASA + 2 g/ ml MC2RR, : MS MC2 RR ASA, 2 g/ ml 2 mmol/ L 3, 112, 1. 4 ROS DCFH2DA [9 ] ; MDA TBA [10 ] ; SOD Prochazko2 va [11 ] [12 ] ; POD [12 ] ;ASA [13 ] ; GSH Griffith [14 ] [12 ] ; G2 250, (BSA) 1. 5 SPSS One2Way ANO2 VA ROS 1,, ROS, ROS 0 4d,,MC - RR ROS, MC - RR + DMSO MC - RR + ASA ROS 1 ROS Fig11 Effects of different treatments on ROS content in tobacco 2. 2 MDA 2, 0 3d, MDA,,MC - RR MDA,MC - RR + DMSO MC - RR + ASA MDA, MC - RR MDA,MC - RR MDA MC - RR + DMSO MC - RR + ASA MDA 2 MDA Fig12 Effects of different treatments on MDA content in tobacco 2. 3 SOD 3, 0 5d,,MC2RR SOD

3 6 : DMSO ASA ROS 871,MC2RR + ASA MC2RR + DMSO SOD, MC2RR SOD, ;MC2RR + DMSO MC2RR + ASA GSH ( ), MC2RR GSH 3 SOD Fig13 Effects of different treatments on SOD activity in tobacco 6,,MC2RR ASA 2. 4 POD, ;MC2RR + ASA,MC2RR POD ASA ;MC2RR + DMSO, ( 4) ASA, MC2RR + DMSO MC2RR + ASA POD MC2RR ASA 0 5d,,,, POD MC2RR 5 GSH Fig15 Effects of different treatments on GSH content in tobacco 2. 6 ASA 6 ASA Fig16 Effects of different treatments on ASA content in tobacco 4 POD Fig14 Effects of different treatments on POD activity in tobacco 2. 5 GSH 5, GSH GSH,MC2RR GSH 3, ROS,,ROS, ROS, [15 ],,ROS,,, ROS,ROS

4 872 31, ROS,MC2 RR ROS,ROS ROS, DNA MC2RR + DMSO MC2RR + ASA ROS, MC2RR, ASA DMSO ROS, ROS, MC2RR, DNA, [16 ] MDA,MC2RR MDA ROS, Ding [17 ],MC ROS, ROS, MDA MC2RR + DMSO MC2RR + ASA MDA, ASA DMSO MC2RR ROS,, SOD POD GSH ASA SOD O 2 - H 2 O 2 1 O 2,POD H 2 O 2 O 2 - H 2 O 2 H 2 O O 2, H 2 O 2,,MC2RR SOD POD,,,,,,SOD POD, POD SOD MC2RR + DMSO MC2RR + ASA SOD POD,, ASA DMSO ROS,,, GSH,, GSH OH HOCl,, GSH GST ASA H 2 O 2, GSH, ROS, GSH ROS MC2RR GSH,, GSH MC2RR ROS MC2RR + DMSO MC2RR + ASA GSH ASA MC2RR,,,, MC2RR, ASA DMSO ROS, ROS MC2RR :,,! : [ 1 ] Yan H, Pan G, Zhang M M. Advances in the study of microcystion toxin [J ]. Acta Ecologica Sinica, 2002,22 (11) : [,,..,2002, 22 (11) : ] [ 2 ] Mackintlsh C,Beattie K A, Klunpp S, et al. Cyanobacterial micro2 cystin2lr is a protein and specific inhibitor of protein phosphatase1 and 2A fron both mammals and higher plants [J ]. FEBS Letters, 1990,264 : [ 3 ] Ding W X,Shen H M,Zhu H G, et al. Hepatotoxicity and formation of reactive oxygen species induced by microcystis aeruginosa toxin [J ]. Chinese Journal of Preventive Medicine,1998,32 (5) : [,,,..,1998,32(5) : ] [ 4 ] Pflugmacher S. Promotion of oxidative stress in the aquatic macrophyte Ceratophyllum demersum during biotransformation of the cyanobacteri2 al toxin microcystin2lr [J ]. Aquatic Toxicology,2004,70(3) : [ 5 ] Yin L Y,Huang J Q,Shen Q, et al. Responses of antioxidant systems in tobacco to the toxicity of microcystin2rr [J ]. China Environmental Science,2005,25 (5) : [,,,..,2005,25(5) : ] [ 6 ] Hu Z Q,Liu Y D. Accumulation of microcystin2rr in Synechococcus elongatus and its toxic effects [ J ]. China Environmental Science, 2005,25 (1) :23 27 [,.

5 6 : DMSO ASA ROS 873.,2005,25 (1) :23 27 ] [ 7 ] Pinho G L L,Moura da Rosa C,Maciel F E, et al. Antioxidant re2 sponses after microcystin exposure in gills of an estuarine crab species pre2treated with vitamin E [ J ]. Ecotoxicology and Environmental Safety,2005,61(3) : [ 8 ] Οegura B, Tamara T L and Filipic M. The role of reactive oxygen species in microcystin2lr2induced DNA damage [ J ]. Toxicology, 2004,200 (1) :59 68 [ 9 ] He Y Y and Hgder D P. UV2B2induced formation of reactive oxygen species and oxidative damage of the cyanobacterium Anabaena sp. : protective effects of ascorbic acid and N2acetyl2L2cysteine [J ]. Jour2 nal of Photochemistry and Photobiology B : Biology, 2002, 66 ( 2) : [10 ] Uchiyama M,Mihara M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test [J ]. Analytical Biochemistry,1978, 86(1) : [11 ] Prochazkova D,Sairam R K,Srivastava G C, et al. Single oxidative stress and antioxidant activity as the basis of senescence in maize leaves [J ]. Plant Science,2001,161(4) : [12 ] Li H S. Principles and techniques of plant physiological biochemical experiment [ M]. Beijing : Higher Education Press. 2000, , [.. :. 2000, , ] [13 ] Chen J X,Wang X F. Guides of plant physiological experiment [M]. Guang Zhou : South China University of Technology Press. 2002, [,.. :. 2002, ] [ 14 ] Griffith O. Determination of glutathione and glutathione disulfide using glutathione reductase and 22vinylpyridine [ J ]. Analytical Biochem2 istry,1980,106 (1) : [15 ] Neill S J,Desikan R,Clarke A, et al. Hydrogen peroxide and nitric oxide as signaling molecules in plants [J ]. Journal of Experimental Botany,2002,53 : [16 ] Ferrat L, Pergent2Martini C, Rom o M. Assessment of the use of biomarkers in aquatic plants for the evaluation of environmental quali2 ty:applications to seagrasses [J ]. Aquatic Toxicology,2003,65 (2) : [17 ] Ding W X, Ong C N. Role of oxidative stress and mitochondrial changes in cyanobacteria2induced apoptosis and hepatotoxicity [J ]. FEMS Microbiology Letters,2003,220(1) :1 7 EFFECTS OF EXO GENOUS DMSO AND ASA ON FORMATION OF REACTIVE OXY GEN SPECIES AND ANTIOXIDANT SYSTEMS OF TOBACCO SUSPENSION CELLS UNDER MC2RR STRESS HUANG Wen2Min 1,2,XING Wei 3,LI Dun2Hai 1 and LIU Yong2Ding 1 (11 Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan ; 21 Graduate School of Chinese Academy of Sciences, Beijing ; 31Wuhan Botanical Garden, the Chinese Academy of Sciences, Wuhan ) Abstract :Microcystins are a family of toxins produced by freshwater cyanobacteria1they are cyclic heptapeptides,composed of sev2 en amino acids1they exert a toxic effect by being specific inhibitors of the protein phosphatases 1 and 2A ( PP1 and PP2A),but re2 cent evidences also indicate its potential to generate oxidative stress1oxidative stress is a general term used to describe the steady state level of oxidative damage in cells,tissues,or organs,caused by the reactive oxygen species ( ROS) 1In this experiment,the toxic mechanism of MC2RR inducing oxidative stress was evaluated in tobacco 1Changes of ROS formation and an2 tioxidant systemof tobacco were studied under the treatment of 2 g/ ml MC2RR,2 g/ ml MC2RR % DMSO and 2 g/ ml MC2RR + 2 mmol/ L ASA1Under the treatment of 2 g/ ml MC2RR,the contents of ROS,MDA and ASA as well as the activities of SOD and POD were higher than those in control after 5 days treatment1gsh content first decreased and then increased,and at the 6 th day of treatment GSH content significantly increased compared with the control1the abundant accumulation of ROS and the change of antioxidant system both indicated that the cells suffered from oxidative stress induced by MC2RR1ROS may play an important role in its toxic effect on the cells1the MC2RR2induced oxidative stress in cells was further confirmed by ex2 posing the cells to MC2RR in the presence of two ROS scavengers,asa or DMSO1ASA has significant capacity of scavenging O2 - and H 2 O 2,and DMSO is a kind of hydroxyl radical scavengers1when tobacco were exposed to 2 g/ ml MC2 RR with 015 % DMSO or 2 mmol/ L ASA,the formation of ROS and MDA were prevented and the contents of GSH,ASA and activi2 ties of SOD and POD all decreased in contrast to MC2RR treatment1it concluded that tobacco suffered oxida2 tive stress after MC2RR treatment and the exogenous ASA and DMSO can protect the cells from MC2RR induced oxidative stress1 Key words :Microcystin2RR ; Tobacco ; Reactive oxygen species ; Antioxidant system; ASA ; DMSO