2014 9 3 517-523 Asian Journal of Ecotoxicology Vol. 9, 2014 No.3, 517-523 DOI: 10.7524/AJE.1673-5897.20140120001. - cry1ab/ac 2014, 9(3): 517-523 Tan S H, Liu Y F, Sun Y D, et al. The response of antioxidant system and metabolic enzyme of Cipangopaludina cathayensis to transgenic Bt gene rice [J]. Asian Journal of Ecotoxicology, 2014, 9(3): 517-523 (in Chinese) - cry1ab /Ac 谭 树 华 1, 刘 雨 芳 1,*, 孙 远 东 1, 肖 璐 1, 李 菲 1, 桂 芳 艳 1, 刘 文 海 1 2, 戈 峰 1. 411201 2. 100101 2014-01-20 2014-02-26 Bt 1 (HH1) Bt 63(MH63) 63(MH63C)3 (SOD CAT GSH-PX GSH) (GST ACP) HH1 SOD CAT GSH-PX MH63 SOD CAT (p < 0.01) GSH GST HH1 MH63 HH1 ACP MH63 (p < 0.01) MH63C MH63 MH63C SOD CAT GST GSH-PX GST GSH MH63C Bt 1673-5897 2014 3-517-07 X171.5 A The Response of Antioxidant System and Metabolic enzyme of Cipangopaludina cathayensis to Transgenic Bt Gene Rice Tan Shuhua 1, Liu Yufang 1,*, Sun Yuandong 1, Xiao Lu 1, Li Fei 1, Gui Fangyan 1, Liu Wenhai 1, Ge Feng 2 1. School of Life Sciences, The Key Laboratory of Horticultural Crops Pest Management in Hunan Province, Huan University of Science and Technology, Xiangtan 411201, China 2. Institute of Zoology, Chinese Academy of Science, Beijng 100101, China Received 20 January 2014 accepted 26 February 2014 Abstract The activities of antioxidative (SOD, CAT, GSH-Px, GSH) and metabolic enzymes (GST, ACP) of Cipangopaludina cathayensis that grown in three kinds of rice habitat including insect-resistant transgenic Bt rice Huahui No 1 HH1, non-bt conventional rice Minghui63 " (MH63 ) without pesticides, and non-bt rice (MH63C) with pesticide were detected in this study. The results showed that the activities of SOD, CAT, GSH- Px of livers and gills of C. cathayensis grown in HH1 rice habitat were all higher than that in MH63 habitat. The difference of SOD and CAT activities between HH1 and MH63 habitat were significant (p < 0.01), while GSH (2012ZX08011002) ( 1972-) E-mail: hstan@126.com * (Corresponding author) E-mail: yfliu2011@126.com
518 9 content and GST activities of livers of C. cathayensis showed no significant difference between HH1 and MH63 rice habitat. The activities of ACP in HH1 rice habitat were higher than in MH63 rice habitat significantly (p < 0.01). As for MH63 and MH63C habitat, the activities of SOD, CAT and GST of livers in MH63C habitat increased significantly, meanwhile GSH-Px GST activities and GSH content of gills in MH63C habitat were activated significantly. It suggested that the antioxidative enzymes and detoxification metabolic enzymes of C. cathayensis under pesticides were activated significantly, while antioxidative enzymes and metabolic enzymes were induced in HH1 transgenic Bt rice habitat. It could be concluded that the toxic effect of Bt rice on C. cathayensis was apparently lower than that of the pesticides. Keywords Cipangopaludina cathayensis; biotech rice; antioxidative enzyme activities; detoxification metabolic enzyme enzymes -, (Bacillus thringiensis (Bt 1 Materials and methods ) Bt 1.1 50 Bt [1] Bt Bt cry1ab/ac 1 [2-5] [6] (HH1) 63(MH63) [7,8] cry1ab/cry1ac [9,10] Bt 1 (HH1) ( 2012 6 9 (2009) 072 ) HH1 1 530 m 2, 9 170 m 2, [11] (Folsomia candida) 3 3 (3 ) 1 [12] 1 (HH1) [13] Bt 2 63 1 (MH63) 1 (MH63C) (Cipangopaludina cathayensis) MH63C 3 7 2 7 21 2% 35% 27% 5 s 25% 8 12 2% 25% 30% 25%,, [14,15], [16] [17] 2.5 g CAT [18] 3 ( = 50 cm 100 cm) 8 m
3 - cry1ab/ac 519 20 cm MH63 (p < 0.01) HH1 CAT 20 10 d MH63C HH1 MH63C CAT MH63 1.2 HH1 MH63C CAT 5 2.513 1.357 0.86% 10% (SOD) (CAT) (ACP) -S (GST) (GSH) (GSH-PX) 1.3 SOD [19],CAT [20] [20],ACP SOD CAT ACP U mg -1 progst CDNB [21] 1 SOD GSH GSH-PX Fig. 1 Comparision the activities of SOD of, Cipangopaludina cathayensis in different rice fields [22], mg ml -1 1.4 SPSS17.0 (ANOVA) S-N-K LSD 2 Results 2.1 SOD Bt 1 (HH1) Bt 63(MH63) Bt 63(MH63C)3 2 SOD 1 HH1 SOD MH63 HH1 SOD MH63 Fig. 2 Comparision the activities of CAT of MH63C SOD HH1 MH63 (p < 0.01) MH63C MH63 (p < 0.05) HH1 Note: ** ** p < 0. 01, p < 0. 01, Same for below CAT 2.3 GSH-Px GSH-Px HH1 MH63 MH63C SOD MH63 111.95% MH63C (p >0.05) GSH-Px MH63 41. 40% HH1 MH63C HH1 MH63C MH63C (p > 0.05) HH1 MH63 (p <0. 01) HH1 MH63 SOD SOD GSH-Px (p >0.05)( 3) MH63C HH1 MH63 MH63 2.4 MH63C HH1 MH63C HH1 MH63 GSH GSH 4 HH1 MH63C (p > 0.05) GSH MH63 2.2 CAT (p > 0.05) MH63 MH63C 3 CAT GSH MH63 GSH 1.69 2 CAT HH1 (p <0.05) HH1 GSH MH63C MH63C SOD MH63
520 9 (p > 0.05)( 6) 3 GSH-Px Fig. 3 The activities of GSH-Px of MH63 HH1 (p < 0.01) MH63C MH63 GST 66.16% HH1 MH63 (p > 0.05) GST GST MH63C MH63C GST HH1 MH63 1.7 2.02 (p < 0.01) HH1 GST MH63 4 GSH * p < 0. 05 Fig. 4 The GSH content of Note: * p < 0. 05 2.5 ACP [23] Cry1Ab/Cry1Ac HH1 ACP Cnaphalocrocis medinalis (Guenée) MH63 MH63C (p < 0.01)HH1 Bt ACP MH63 MH63C SOD CAT POD 3 112.4% 98.8% MH63C MH63 ACP MH63C HH1 MH63 3 ACP Cry1Ab/Cry1Ac 1 (HH1) (p > 0.05)( 5) 5 ACP Fig. 5 The activities of ACP of 2.6 GST 6 GST Fig. 6 The activities of GST of 3 Discussion MH63 HH1 SOD CAT GSH- PX MH63 SOD CAT 3 O - H 2 2 O 2 HH1 Bt O - H 2 2 O 2 Bt Bt Bt Bt Bt MH63C GST
3 - cry1ab/ac 521 GSH GST GSH GSH-Px GST GST Bt Bt GSH, [29] Bt GSH, (Se-GPx) [30] [24] [25] Bt Cry1Ab 63 [31-34] Bt Bt, [35] HH1 MH63 Bt GSH GST 刘 雨 芳 ( 1964 ), 女, 博 士, 教 授, 硕 士 生 导 GST GSH HH1 师 主 要 研 究 方 向 为 农 业 生 态 学 环 境 生 态 安 全 评 价, 发 表 论 文 50 多 篇 Bt ACP [ 1 ],,,. Bt [J]., 2005, 19(1): [26] [27] 74-78 Bt Zhou X, Cheng J A, Hu Y, et al. Effects of transgenic Bt rice on the population development of Nephotettix cincticeps [J]. Chinese Journal of Rice Science, 2005, 19(1): 74 11 HH1 ACP - 78 (in Chinese) MH63 HH1 [ 2 ],,,. DNA 2005, 38(4): 725-729 Liu Y F, Wang F, You M S, et al. Resistance of cry1ac + SCK transgenic rice and its filial generation to the rice leaf roller Cnaphalocrocis medinalis [ J ]. Scientia Agricultura HH1 Sinica, 2005, 38(4): 725-729 (in Chinese) [ 3 ],,,. Bt MH63 HH1 2008, 51(10): 1022-1027 Zhang W, Zhang Z G, Fu X Q, et al. Effects of transgenic Bt MH63C rice on the activities of three protective enzymes in larvae of the rice leaffolder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) [J]. Acta Entomologica Sinica, 2008, 51 HH1 Bt (10): 1022-1027 (in Chinese) Bt [ 4 ] Ye G Y, Shu Q Y, Yao H W, et al. Field evaluation of Bt resistance of transgenic rice containing a synthetic cryab [28] MH63C MH63 gene from Bacillus thuringiensis Berliner to two stem ACP borers [J]. Journal of Economic Entomology, 2001, 94: 271-276
522 9 [ 5 ] Khanna H K, Raina S K. Elite Indica transgenic rice plants expressing modified Cry1Ac endotoxin of Bacillus thuringiensis show enhanced resistance to yellow stem borer (Scirpophaga incertulas) [J]. Transgenic Research, 2002, 11: 411 transgenic rice to the rhizosphere microorganisms in the Plant Breeding Base of Hainan [J]. China journal of biosafety 2012, 21(1): 61-66 (in Chinese) [14] Livingstone P R. Accumulation of heavy metals in invertebrates and its ecological aspects [ J]. Comparative Biochemistry & Physiology, 1991, 100: 151-155. [15] Burbidge F J, Jacey D J, Webb J, et al. A comparison between particulate (elemental) zinc and soluble zinc(zncl 2 ) - 423 [ 6 ] Liu W, Hao L H, Wu W, et al. Transgenic Bt rice does not affect enzyme activities and microbial composition in the rhizosphere during crop development [J]. Soil Biology Biochemistry, 2008, 40(2): 475-486 uptake and effects in the mussel, Mytilusedulis [ J]. Archives [ 7 ] Bai Y Y, Yan R H, Ye G Y, et al. Field response of aboveground non-target arthropod community to transgenic of Environmental Contamination. Toxicology, 1994, 26: 446-472 Bt-Cry1Ab rice plant residues in postharvest seasons [J]. Transgenic Research, 2012, 21: 1023-1032 [16] Walsh K, Dunstan R H, Murdoch R N, et al. Bioaccumulation of pollutants and changes in population parameters in [ 8 ],,,. cry1ac/sck the Gastropod Mollus Austocochlea Constrica [ J ]. Archives [J]. 2006 49(6): 955-962 Liu Y F, He L, Wang Q, et al. Evaluation of the effects of insect-resistant cry1ac/scktransgenic rice on the parasitoid communities in paddy fields [J]. Acta Entomologica Sinica, 2006, 49(6): 955-962 (in Chinese) [17] of Environmental Contamination. Toxicology, 1994, 26: 367-373,. 1997, 12(2): 8-11 Guo M X, Lin R H. River Snail Cipangopaludina cathayensis [ 9 ] as an indicator for toxicity and bioavailability of heavy,,,. Bt [J]., 2011, 54(4): 467-476 Gao M Q, Hou S P, Pu D Q, et al. Effects of Bt rice on the number and hatch rate of planthopper eggs and their attack by natural enemies in paddy fields [J]. Acta Entomologica Sinica, 2011, 54(4): 467-476 (in Chinese) [18] metals in sediment [ J ]. Environment and Development, 1997, 12(2): 8-11 (in Chinese),,,. [J]., 2012, 27(4): 37-40 Lei Y L, Yin W Z M, Zhang Y, et al. Effect of cadmium [10],,,. cry1ac/sck and mercury on catalase activity of the mudsnail [J]. Journal of Neijiang Normal University, 2012, 27(4): 37-40 (in [J]., 2007, 40(6): 1181-1189 Liu Y F, He L, Wang Q, et al. Effects of and ecological safety insect-resistant Cry1Ac/sck transgenic rice on key non-target pests in paddy fields [ J]. Scientia Agricultura Sinica, 2007, 40(6): 1181-1189 (in Chinese) [19] Chinese),,,. SOD [J]., 1985, 13(4): 71-73 Zou G L, Gui X F, Zhong X L, et al. Determination method [11] Li Y Z, Sui H, Chen F J, et al. Effects of insect-resistant transgenic Bt rice with a fused Cry1Ab + Cry1Ac gene on population dynamics of the stem borers, Chilosuppressalis of SOD pyrogllol autoxidation improvement [J]. Progress in Biochemistry and Biophysics, 1986, 13 (4): 71-73 (in Chinese) and Sesamiainferens, occurring in paddy field [ J ]. Acta Ecologica Sinica, 2012, 32(6): 1783-1789 [20]. [M].,, 1985, 36-37, 62-64 [12] Yuan Y Y, Xiao N W, Ge F, et al. Laboratory assessment of the impacts of transgenic Bt rice on the ecological fitness of the soil non-target arthropod, Folsomia candida (Collembola: Isotomidae) [J]. Transgenic Research, 2013, 22: 791 [21] Luca-Abbott S B D, Richardson B J, McClellan K E, et al. Field validation of antioxidant enzyme biomarkers in mussels (Pernaviridis) and clams (Ruditapesphilip- pinarum) transplanted in Hong Kong coastal waters [J]. Marine Envi- - 803 ronment Research, 2005, 51: 694-707 [13],,,. [22] Bradford M M. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the [J]., 2012, 21(1): 61-66 principle of protein-dye binding [J]. Analytical Biochemistry, 1976, 72: 248 - Xu L, Huang Q X, Zou J, et al. Study of the effects of 254
3 - cry1ab/ac 523 [23],,,. Bt assessing genetically engineered insect-resistance in trees. Agriculture and Human Values, 1997, 14(3): 237-249 2008, 51(10): 1022-1027 Zhang W, Zhang Z G, Fu X Q, et al. Effects of transgenic Bt rice on the activities of three protective enzymes in larvae of the rice leaffolder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae) [J]. Acta Entomologica Sinica, 2008, 51 (10): 1022-1027 (in Chinese) [30] [31] Saxena D, Flores S, Stotzky G. Vertical movement in soil of insecticidal Cry1Ab protein from Bacillus thuringiensis [J]. Soil Biology Biochemistry, 2002, 34: 111-120,,,. Bt Cry1Ab., 2004, 18(3): 255-261 [24]. [J]. Bai Y Y, Jiang M X, Cheng J A, et al. Degradation of, 1994, 21(5): 395-399 Chen Y K. Detoxication and toxic metabolite of glutathione [J]. Progress in biochemistry and Biophysics, 1994, 21(5): 395-399 [32] Cry1Ab toxin protein expressed by Bt transgenic rice in paddy soils. Chinese Journal Rice Science, 2004, 18 (3): 255-261 (in Chinese),,,. Bt [25],,,. Bt (Pardosa Psoudoannulata) [J]., 2011, 20(1): 45-49 Liu L J, Huang Y, duan Y H, et al. Effects of Bt protein expressed by transgenic rice on the development of wolf spider (Parddsa pseudoannulata) [J]. Acta Laser Biology Sinica, 2011, 20(1): 45-49 (in Chinese) 2011, 50 (24): 5200-5205 Xiong Y F, Qin T C, Chen C, et al. Effects of transgenic Bt rice on the activities of protective enzymes and detoxification enzymes in Kunming male mouse [J]. Hubei Agricultural Sciences. 2011, 50(24): 5200-5205 (in Chinese) [33],,,. Bt [26],,,. V E [J]., 2012, 32 (11): 3546 -., 3554 2002 21(4): 431-438 Ai C X, Chen L Q, Wen X B, et al. Effect of vitamin E on SOD, ALP and ACP activities of Chinese mitten-handed crab, Eriocheir sinensis [ J ]. Journal of Oceanography in Taiwan Strait, 2003, 22(1): 24-31 (in Chinese) [34] Zhu X Y, Li Z Y, Chang L, et al. Community structure and abundance dynamics of soil collembolans in transgenic Bt rice paddy fields [J]. Acta Ecologica Sinica, 2012, 32(11): 3546-3554 (in Chinese),,,. Bt 63 [27],,,. Bt [J]., 2008, 4: 31-33 Yu C, Zhang J, Ye S H, et al. A preliminary study on the physiological and biochemical characteristics of Bt rice [J]. Jiangsu Agricultural Sciences, 2008, 4: 31-33 (in Chinese) 2012, 23(11): 3065-3071 Li X Q, Chen F J, Liu M Q, et al. Effects of two years planting transgenic Bt rice (BtSY63) on soil nematode community [J]. Chinese Journal of Applied Ecology, 2012, 23 [28],,. (11): 3065-3071 (in Chinese) 2004, [35] 15(12): 2309-2314 Liu Z C, Ye G Y, Hu C, et al. Effects of Bacillus thuringiensis transgenic rice and chemical insecticides on arthropod communities in paddy-fields [J]. Chinese Journal of Applied Ecology, 2004, 15(12): 2309-2314 (in Chinese),,. Bt Cry1Ab 2005, 25(7): 1583-1590 Temporal expression patterns of Cry1Ab insecticidal protein in Bt rice plants and its degradation in paddy soils [J]. Acta Ecologica Sinica, 2005, 25(7): 1583-1590 (in Chinese) [29] James R. Utilizing a social ethic toward the environment in