转基因抗虫棉Bt毒蛋白含量时空变化及土壤降解研究Study on Spatial-temporal Dynamics of Bt Toxic Protein Expression in Insect-resistant Transgenic Cotton and Its Degradation in Soil
张益文;王连荣;张连成;张军;籍新波;王进茂;
摘要(Abstract):
[目的]研究转基因抗虫棉Bt毒蛋白含量的时空变化及其土壤降解。[方法]采用ELISA法(酶链免疫法)研究和分析了转Bt-cry1Ac基因抗虫棉的根、茎和叶片组织在不同发育时期毒蛋白的含量变化及转Bt基因抗虫棉(GK45)和非转基因棉花(新陆早36号)在根际土壤、表层土壤和后茬种植区土壤中Btcry1Ac毒蛋白的年平均含量变化。[结果]BtCry1Ac毒蛋白含量在抗虫棉生长过程中均呈动态下降趋势,而根中下降的速率最快,茎和叶片次之;棉花种植区土壤表层中均检测到Btcry1Ac毒蛋白,且后茬种植区中表层毒蛋白的含量增加,而根际土中含量极低。[结论]Btcry1Ac毒蛋白的含量检测为种植转基因作物的风险评价及转基因作物的土壤生态系统安全性评价提供了科学依据。
关键词(KeyWords): 转基因棉花;Bt毒蛋白;土壤微生物;安全性评价
基金项目(Foundation): 转基因生物新品种培育重大专项(2009ZX08011-027B);; 国家自然科学基金资助项目(30872050)
作者(Author): 张益文;王连荣;张连成;张军;籍新波;王进茂;
Email:
DOI: 10.13989/j.cnki.0517-6611.2012.25.096
参考文献(References):
- [1]YANG L,CHEN J,HUANG C,et al.Validation of a Cotton-Specific Gene,Sad1,Used as an Endogenous Reference Gene in Qualitative and Real-Time Quantitative Pcr Detection of Transgenic Cottons[J].Plant Cell Re-ports,2005,24(4):237-245.
- [2]TABASHNIK B E,GASSMANN A J,CROWDER D W,et al.Insect Resist-ance to Bt Crops:Evidence Versus Theory[J].Nat Biotech,2008,26(2):199-202.
- [3]邢珍娟,王振营,何康来,等.转Bt基因抗虫玉米根茬和根际土壤中BtCry1ab杀虫蛋白的田间降解动态[J].中国农业科学,2010(23):4970-4976.
- [4]SABHARWAL N,ICOZ I,SAXENA D,et al.Release of the RecombinantProteins,Human Serum Albumin,Beta-Glucuronidase,Glycoprotein B fromHuman Cytomegalovirus,and Green Fluorescent Protein,in Root Exudatesfrom Transgenic Tobacco and Their Effects on Microbes and Enzymatic Ac-tivities in Soil[J].Plant Physiol Biochem,2007,45(6/7):464-469.
- [5]ICOZ I,STOTZKY G.Cry3bb1 Protein from Bacillus Thuringiensis in RootExudates and Biomass of Transgenic Corn Does Not Persist in Soil[J].Transgenic Res,2008,17(4):609-620.
- [6]YUAN Y G,GE F.Effects of Transgenic Bt Crops on Non-Target Soil Ani-mals[J].Ying Yong Sheng Tai Xue Bao,2010,21(5):1339-1345.
- [7]ZHOU X Y,LIU N,ZHAO M,et al.Advances in Effects of InsecticidalCrystal Proteins Released from Transgenic Bt Crops on Soil Ecology[J].YiChuan,2011,33(5):443-448.
- [8]SAXENA D,STOTZKY G.Bacillus Thuringiensis(Bt)Toxin Releasedfrom Root Exudates and Biomass of Bt Corn Has No Apparent Effect onEarthworms,Nematodes,Protozoa,Bacteria,and Fungi in Soil[J].Soil Bi-ology and Biochemistry,2001,33(9):1225-1230.
- [9]SAXENA D,STOTZKY G.Bt toxin is not taken up from soil or hydroponicculture by corn,carrot,radish,or turnip[J].Plant and Soil,2002,239:165-172.
- [10]ICOZ I,SAXENA D,ANDOW D A,et al.Microbial Populations and En-zyme Activities in Soil in Situ under Transgenic Corn Expressing Cry Pro-teins from Bacillus Thuringiensis[J].J Environ Qual,2008,37(2):647-662.
- [11]安鑫龙,董金皋,韩建民.玉米大斑病菌的rapd分析:I应用CTAB法提取玉米大斑病菌DNA[J].河北农业大学学报,2001(1):38-41.
- [12]夏兰芹,徐琼芳,郭三堆.抗虫棉生长发育过程中Bt杀虫基因及其表达的变化[J].作物学报,2005(2):197-202.
- [13]孙彩霞,陈利军,武志杰,等.种植转Bt基因水稻对土壤酶活性的影响[J].应用生态学报,2003(12):2261-2264.
- [14]SANDER M,MADLIGER M,SCHWARZENBACH R P.Adsorption ofTransgenic Insecticidal Cry1ab Protein to SiO2.1.Forces Driving Adsorp-tion[J].Environ Sci Technol,2010,44(23):8870-8876.
- [15]MADLIGER M,GASSER C A,SCHWARZENBACH R P,et al.Adsorptionof Transgenic Insecticidal Cry1ab Protein to Silica Particles.Effects onTransport and Bioactivity[J].Environ Sci Technol,2011,45(10):4377-4384.
- [16]张永军,吴孔明,郭予元.转Bt基因棉花杀虫蛋白含量的时空表达及对棉铃虫的毒杀效果[J].植物保护学报,2001(1):1-6.
- [17]孙彩霞,陈利军,武志杰.Bt毒素在转基因棉花与土壤系统中的分布[J].应用生态学报,2005(9):1765-1768.
- [18]SAXENA D,FLORES S,STOTZKY G.Insecticidal Toxin in Root Exu-dates from Bt Corn[J].Nature,1999,402(6761):480.
- [19]芮玉奎.Bt棉与常规棉根际土壤Bt毒蛋白和植物激素变化动态[J].生物技术通讯,2005(5):515-517.
- [20]沈平,张永军,陈洋,等.Bt棉不同种植年限土壤中Bt基因及其蛋白的残留测定[J].棉花学报,2008(1):79-81.
- [21]HEAD G,SURBER J B,WATSON J A,et al.Nodetection of Cry1ac Pro-tein in Soil after Multiple Years of Transgenic Bt Cotton(Bollgard)Use[J].Environmental Entomology,2002,31:30-36.
- [22]XUE L,LU Y,ZHOU Z T,et al.Effect of transgenic Bt plus CpTI cottonon carboxylesterase and acetylcholinesterase of cotton aphid Aphis gossy-pii[J].Agricultural Science&Technology,2011,12(7):997-1000.
- [23]金黎明,李力,文泽会,等.转Bt基因抗虫杂交棉产量与重要性状的多元线性回归分析[J].安徽农业科学,2010,38(17):8894-8896,8899.