转mCry1Ac基因玉米BT799对斑马鱼的生态毒理学效应
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摘要
转基因作物的饲用安全性问题一直是人们关注的焦点之一.为评估转mCry1Ac基因玉米BT799对鱼类的生态毒理效应,本研究以斑马鱼为受试动物,设置5个处理:含20%转基因玉米膨化饲料组(GMF)、含20%亲本玉米膨化饲料组(PF)、转基因玉米粉(GMM)、亲本玉米粉(PMM)以及商业饲料对照组(CF),通过98 d的喂养试验,调查斑马鱼的生长表现、组织病理、繁殖、肝脏中抗氧化酶活性及敏感蛋白mRNA的表达水平.结果表明:转mCry1Ac基因玉米对斑马鱼的各项生长指标、肝脏、脑和肠道的组织病理、产卵量、受精卵孵化率、肝脏中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性,以及SOD、CAT、热激蛋白70(HSP70)和卵黄蛋白原(VTG)mRNA表达量均无显著影响.但在试验后期,饲料组(GMF和PF)和玉米粉组(GMM和PMM)斑马鱼的体重、体长和特定生长率显著低于商业饲料组;饲料组斑马鱼的孵化率显著低于玉米粉组和商业饲料组;饲料组(3.85±0.76)雄鱼肝脏中的VTG mRNA表达量显著高于玉米粉组(1.60±0.56).研究表明,转mCry1Ac基因玉米对斑马鱼没有明显生态毒理效应,但由于配制的膨化饲料与商业饲料在营养成分和适口性上的差异,可能导致个别指标与商业饲料组相比有显著差异.
The safety of feed derived from genetically modified(GM) crops is one of the focuses of attention. To evaluate the ecotoxicological effects of transgenic mCry1Ac maize(BT799) on fish, zebrafish(Danio rerio) were fed extruded feeds containing either 20% GM maize(GMF) or its parental control maize(PF), GM maize meal(GMM) or its parental control maize meal(PMM), and a control commercial feed(CF), respectively. The growth performance, histopathology, reproduction, antioxidant enzyme activity and mRNA expression levels of sensitive protein in the liver were investigated over the course of a 98-day feeding trial. The results showed that transgenic mCry1Ac maize had no significant effect on growth, histopathology of the liver, brain and intestinal tract, fecundity, hatching rate of fertilized eggs, superoxide dismutase(SOD), catalase(CAT) activity, mRNA expression levels of SOD and CAT, or heat shock protein 70(HSP70) and vitellogenin(VTG) in the liver. However, zebrafish fed the commercial feed exhibited significantly greater weight, longer length, and higher specific growth rate than those fed feeds(GMF and PF) and maize meals(GMM and PMM). The hatching rate of zebrafish in the feed groups was significantly lower than that of the maize meal groups and the commercial feed group. The mRNA transcriptional levels of VTG were significantly higher in the liver for the feed groups(3.85±0.76) than that for the maize meal groups(1.60±0.56). These results suggest that transgenic mCry1Ac maize has no ecotoxicological effects on zebrafish. However, the differences in nutrient composition and palatability between the extruded experimental feeds and the commercial feed would lead to significant diffe-rences in some parameters.
The safety of feed derived from genetically modified(GM) crops is one of the focuses of attention. To evaluate the ecotoxicological effects of transgenic mCry1Ac maize(BT799) on fish, zebrafish(Danio rerio) were fed extruded feeds containing either 20% GM maize(GMF) or its parental control maize(PF), GM maize meal(GMM) or its parental control maize meal(PMM), and a control commercial feed(CF), respectively. The growth performance, histopathology, reproduction, antioxidant enzyme activity and mRNA expression levels of sensitive protein in the liver were investigated over the course of a 98-day feeding trial. The results showed that transgenic mCry1Ac maize had no significant effect on growth, histopathology of the liver, brain and intestinal tract, fecundity, hatching rate of fertilized eggs, superoxide dismutase(SOD), catalase(CAT) activity, mRNA expression levels of SOD and CAT, or heat shock protein 70(HSP70) and vitellogenin(VTG) in the liver. However, zebrafish fed the commercial feed exhibited significantly greater weight, longer length, and higher specific growth rate than those fed feeds(GMF and PF) and maize meals(GMM and PMM). The hatching rate of zebrafish in the feed groups was significantly lower than that of the maize meal groups and the commercial feed group. The mRNA transcriptional levels of VTG were significantly higher in the liver for the feed groups(3.85±0.76) than that for the maize meal groups(1.60±0.56). These results suggest that transgenic mCry1Ac maize has no ecotoxicological effects on zebrafish. However, the differences in nutrient composition and palatability between the extruded experimental feeds and the commercial feed would lead to significant diffe-rences in some parameters.
引文
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[24] Sanden M,Krogdahl ?,Bakke-Mckellep AM,et al.Growth performance and organ development in Atlantic salmon,Salmo salar L.parr fed genetically modified (GM) soybean and maize.Aquaculture Nutrition,2006,1:1-14
[25] Goolish EM,Okutake K,Lesure S.Growth and survivorship of larval zebrafish Danio rerio on processed diets.North American Journal of Aquaculture,1999,61:189-198
[26] Fr?ystad MK,Lilleeng E,Bakke-McKellep AM,et al.Distal intestinal gene expression in Atlantic salmon (Salmo salar L.) fed genetically modified maize.Aquaculture Nutrition,2009,15:104-115
[27] Sissener NH,Bakke AM,Gu J,et al.An assessment of organ and intestinal histomorphology and cellular stress response in Atlantic salmon (Salmo salar L.) fed genetically modified Roundup Ready? soy.Aquaculture,2009,298:101-110
[28] Sagstad A,Sanden M,Krogdahl ?,et al.Organs deve-lopment,gene expression and health of Atlantic salmon (Salmo salar L.) fed genetically modified soybeans compared to the near-isogenic non-modified parental line.Aquaculture Nutrition,2008,14:556-572
[29] L?nge R,Hutchinson TH,Croudace CP,et al.Effects of the synthetic estrogen 17α-ethinylestradiol on the life-cycle of the fathead minnow (Pimephales promelas).Environmental Toxicology and Chemistry,2001,20:1216-1227
[30] Bennetau Pelissero C,Breton B,Bennetau B,et al.Effect of genistein-enriched diets on the endocrine process of gametogenesis and on reproduction efficiency of the rainbow trout Oncorhynchus mykiss.General and Comparative Endocrinology,2001,121:173-187
[31] Duke SO,Rimando AM,Pace PF,et al.Isoflavone,glyphosate,and aminomethylphosphonic acid levels in seeds of glyphosate-treated,glyphosate-resistant soybean.Journal of Agricultural and Food Chemistry,2003,51:340-344
[2] Finamore A,Roselli M,Britti S,et al.Intestinal and peripheral immune response to MON810 maize ingestion in weaning and old mice.Journal of Agricultural and Food Chemistry,2008,56:11533-11539
[3] Prescott VE,Campbell PM,Moore A,et al.Transgenic expression of bean α-amylase inhibitor in peas results in altered structure and immunogenicity.Journal of Agricultural and Food Chemistry,2005,53:9023-9030
[4] Delaney B,Astwood JD,Cunny H,et al.Evaluation of protein safety in the context of agricultural biotechnology.Food and Chemical Toxicology,2008,46:71-97
[5] Marsteller N,B?gh KL,Goodman RE,et al.A review of animal models used to evaluate potential allergenicity of genetically modified organisms (GMOs).Drug Discovery Today:Disease Models,2015,17:81-88
[6] Ronaldw H.Utilization of plant proteins in fish diets:Effects of global demand and supplies of fishmeal.Aquaculture Research,2010,41:770-776
[7] Breitenbach J,Nogueira M,Farré G,et al.Engineered maize as a source of astaxanthin:Processing and application as fish feed.Transgenic Research,2016,25:785-793
[8] Dong S-S (董姗姗),Zhang D-N (章嫡妮),Zhang Z-H (张振华),et al.Overview of ecotoxicological effects of genetically modified crops on fish.Asian Journal of Ecotoxicology (生态毒理学报),2017,12(5):26-34 (in Chinese)
[9] Sissener NH,Sanden M,Krogdahl ?,et al.Genetically modified plants as fish feed ingredients.Canadian Journal of Fisheries and Aquatic Sciences,2011,68:563-574
[10] Sissener NH,Johannessen LE,Hevr?y EM,et al.Zebrafish (Danio rerio) as a model for investigating the safety of GM feed ingredients (soya and maize),performance,stress response and uptake of dietary DNA sequences.British Journal of Nutrition,2010,103:3-15
[11] Cellini F,Chesson A,Colquhoun I,et al.Unintended effects and their detection in genetically modified crops.Food & Chemical Toxicology,2004,42:1089-1125
[12] Somers DA,Makarevitch I.Transgene integration in plants:Poking or patching holes in promiscuous genomes?Current Opinion in Biotechnology,2004,15:126-131
[13] Papst C,Utz HF,Melchinger AE,et al.Mycotoxins produced by Fusarium spp.in isogenic Bt vs.non-Bt maize hybrids under European corn borer pressure.Agronomy Journal,2005,97:219-224
[14] Saxena D,Stotzky G.Bt corn has a higher lignin content than non-Bt corn.American Journal of Botany,2001,88:1704-1706
[15] Hemre GI,Sagstad A,Bakke-Mckellep AM,et al.Nutritional,physiological,and histological responses in Atlantic salmon,Salmo salar L.,fed diets with genetically modfied maize.FEMS Yeast Research,2007,13:186-199
[16] Sagstad A,Sanden M,Haugland ?,et al.Evaluation of stress- and immune-response biomarkers in Atlantic sal-mon,Salmo salar L.,fed different levels of genetically modified maize (Bt maize),compared with its near-isogenic parental line and a commercial suprex maize.Journal of Fish Diseases,2007,30:201-212
[17] NRC.Nutrient Requirement of Fish.Washington,DC:National Academy Press,1993
[18] Gu J,Bakke AM,Valen EC,et al.Bt-maize (MON810) and non-GM soybean meal in diets for Atlantic salmon (Salmo salar L.) juveniles:Impact on survival,growth performance,development,digestive function,and transcriptional expression of intestinal immune and stress responses.PLoS One,2014,9(6):e99932
[19] Chen S (陈松) ,Huang J-Q (黄骏麒),Zhou B-L (周宝良),et al.An assessment on the safety of cottonseed meal from Bt-transgenic cotton plant on the chronic toxicity test with carp.Cotton Science (棉花学报),1996,8(5):241-245 (in Chinese)
[20] Chainark P,Satoh S,Hino T,et al.Availability of genetically modified soybean meal in rainbow trout Oncorhynchus mykiss diets.Fisheries Science,2006,72:1072-1078
[21] Zhu M-X (祝梅香),Wang T-Q (王天奇),Zhang D-Y (张大勇),et al.Nutritional requirements and current status of diets of laboratory zebrafish and swordtails.Chinese Journal of Comparative Medicine (中国比较医学杂志),2009,19(12):61-65 (in Chinese)
[22] Lawrence C.The husbandry of zebrafish (Danio rerio):A review.Aquaculture,2007,269:1-20
[23] Sanden M,Ornsrud R,Sissener NH,et al.Cross-gene-rational feeding of Bt (Bacillus thuringiensis)-maize to zebrafish (Danio rerio) showed no adverse effects on the parental or offspring generations.British Journal of Nutrition,2013,110:2222-2233
[24] Sanden M,Krogdahl ?,Bakke-Mckellep AM,et al.Growth performance and organ development in Atlantic salmon,Salmo salar L.parr fed genetically modified (GM) soybean and maize.Aquaculture Nutrition,2006,1:1-14
[25] Goolish EM,Okutake K,Lesure S.Growth and survivorship of larval zebrafish Danio rerio on processed diets.North American Journal of Aquaculture,1999,61:189-198
[26] Fr?ystad MK,Lilleeng E,Bakke-McKellep AM,et al.Distal intestinal gene expression in Atlantic salmon (Salmo salar L.) fed genetically modified maize.Aquaculture Nutrition,2009,15:104-115
[27] Sissener NH,Bakke AM,Gu J,et al.An assessment of organ and intestinal histomorphology and cellular stress response in Atlantic salmon (Salmo salar L.) fed genetically modified Roundup Ready? soy.Aquaculture,2009,298:101-110
[28] Sagstad A,Sanden M,Krogdahl ?,et al.Organs deve-lopment,gene expression and health of Atlantic salmon (Salmo salar L.) fed genetically modified soybeans compared to the near-isogenic non-modified parental line.Aquaculture Nutrition,2008,14:556-572
[29] L?nge R,Hutchinson TH,Croudace CP,et al.Effects of the synthetic estrogen 17α-ethinylestradiol on the life-cycle of the fathead minnow (Pimephales promelas).Environmental Toxicology and Chemistry,2001,20:1216-1227
[30] Bennetau Pelissero C,Breton B,Bennetau B,et al.Effect of genistein-enriched diets on the endocrine process of gametogenesis and on reproduction efficiency of the rainbow trout Oncorhynchus mykiss.General and Comparative Endocrinology,2001,121:173-187
[31] Duke SO,Rimando AM,Pace PF,et al.Isoflavone,glyphosate,and aminomethylphosphonic acid levels in seeds of glyphosate-treated,glyphosate-resistant soybean.Journal of Agricultural and Food Chemistry,2003,51:340-344